PATENT ABSTRACT
A family of somatotropin vectors with which micro-organisms, preferably bacteria such as, but not limited to  E. coli , can be transformed to enable the expression of bovine somatotropin at high levels using conventional fermentation and induction conditions.

PATENT DESCRIPTION
CROSS-REFERENCE TO RELATED APPLICATIONS  
       [0001]    In accordance with 35 U.S.C. §§119(e) and 365(c) this continuation-in-part application claims the benefit of the filing date of co-pending international application PCT/US01/49977, with an international filing date of Dec. 21, 2001, which claims the benefit of U.S. provisional application No. 60/258,291, filed Dec. 26, 2000, the entirety of each of which is herein incorporated by reference. 
     
    
     
       BACKGROUND OF THE INVENTION  
         [0002]    In mammals, growth hormones (somatotropins) are initially expressed in a pre-hormone form comprising a leader sequence which is removed during secretion of the mature, biologically active hormone from pituitary cells. As used herein, “mature” somatotropin (“ST”) means having an amino acid length essentially like those of the ST secreted into the bloodstream of the aninial, e.g., N-Ala-Phe-Pro- or N-Phe-Pro-bovine ST, N-Phe-Pro-human, -porcine (identical to canine) or -equine ST, etc.  
           [0003]    Attempts to bacterially express human and bovine STs in non-secretion systems using structural genes having the sequences of their cDNAs were at first unsuccessful. Researchers succeeded after introducing silent mutations into the front end of the structural gene. P. H. Seeburg et al., attributed the original difficulties to translation being impeded by secondary structure of the mRNA corresponding to cDNA, and taught lessening such secondary structure to enable significant expression. DNA, Vol. 2, No. 1, 1983, pp. 37-45; U.S. Pat. Nos. 5,254,463 and 5,260,201.  
           [0004]    It has been possible to obtain commercially viable ST expression levels using DNA containing certain silent mutations. However, the need for new vectors that are highly productive and less expensive to use has been difficult to satisfy because the relationships between the configurations and expression levels of “mature” somatotropin vectors remain poorly understood and highly unpredictable.  
         SUMMARY OF THE INVENTION  
         [0005]    Described herein is a family of ST vectors with which micro-organisms, preferably bacteria such as, but not limited to  E. coli , can be transformed to enable the expression of bovine ST (“bST”) at high levels using conventional fermentation and induction conditions. This family is exemplified by the 44 vectors listed in Table I each of which comprises the corresponding sequence of SEQ ID NOs 1 through 44, as indicated in Table I. Each of SEQ ID NOs 1 through 44, extends from an EcoRI site (GAATTC) before the promoter region, through the promoter, ribosome binding site (“RBS”) and entire bST structural gene, ending with the translation stop codon for the structural gene. 
       
    
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0006]    Each of the vectors having SEQ ID NOs 1-44 includes a synthetic promoter, herein designated “cpex-20”, which is disclosed in PCT Appln. PCT/US00/40014 filed Mar. 31, 2000, which claims priority of U.S. Provisional Patent Appln. No. 60/127,449, filed Apr. 1, 1999. This promoter is situated between the EcoRI site (GAATTC) and the AscI site (GGCGCGCC). Various other conventional and novel promoters can be used in these vectors in lieu of cpex-20 to achieve good levels of ST expression.  
         [0007]    To construct the expression vector pXT709, the synthetic EcoRI-HindIII DNA fragment shown in FIG. 1 (SEQ ID NO:50) was ligated into pBR322 (Boehringer Mannheim, Indianapolis, Ind.) which had been digested with EcoRI and HindIII. The resulting plasmid, designated pXT709, was then transformed into recA+  E. coli  K-12 host strain and somatotropin production was induced with nalidixic acid.  
         [0008]    To construct similar plasmids comprising the other sequences listed in the Sequence Listing (namely SEQ ID NOs 1-15, and 17-44), the corresponding synthetic DNA fragments could be similarly prepared and ligated into pBR322. Furthermore, the many other techniques and/or procedures for ligating the described DNA fragments, as shown in the Sequence Listing, into various other plasmids are known and routinely practiced by those of ordinary skill in the art. Moreover, plasmids suitable for transforming  E. coli  and/or other bacteria so as to achieve somatotropin production from cultures of the transformed bacteria are known and regularly used by ordinarily skilled artisans.  
         [0009]    SEQ ID NOs 1-17 have an identical RBS, CATAACATCAAGAGGATATGAAATT (SEQ ID NO:45), herein designated “dps”, that is native to  E. coli.  SEQ ID NOs 18-29 share a novel synthetic RBS, GTACAAATCATAGAGGGTATTTAAT (SEQ ID NO:46) which is herein designated “L49”. SEQ ID NOs 30-42 share a second novel synthetic RBS, AGACACTAAATAGAGGGTATTTAATT (SEQ ID NO:47) herein designated “L437”. Seq. Nos. 43-44 share a third novel synthetic RBS, TAAAAGAGACTAGGAGGAGATTAGA (SEQ ID NO:48), herein designated “R806”. Each RBS is situated between the AscI site and the ATG translation start codon of the bST structural gene. The AscI site begins at coordinate 76 of each sequence. The ATG start codon begins at approximately coordinate 109; this can vary slightly due to the RBSs employed having slightly different lengths.  
         [0010]    All of the vectors listed in Table I (respectively comprising SEQ ID NOS 1-44) have structural genes that differ from cDNA encoding “mature” bST by containing various arrays of silent changes within the nucleotides encoding the first 16 codons of bST. As shown in FIG. 1, vector pXT709 (SEQ ID NO: 50) differs from pXT674 (SEQ ID NO: 49) by having an additional 12 codons within the structural gene changed from those of cDNA, 11 to  E. coli -preferred codons and a twelfth to eliminate an undesirable restriction site, by changing the translation stop codon to two tandem stop codons, and by substituting a different terminator which has two lacUV5 promoters in tandem. pXT703 (comprising SEQ ID NO:15) and pXT747 (comprising SEQ ID NO:17) differ from pXT601 (comprising SEQ ID NO:2) and pXT686 (comprising SEQ ID NO:13), respectively, in the same way.  
         [0011]    Each of the vectors comprising SEQ ID NOs 1 through 44 have expressed mature bST at unoptimized levels of at least about 3.5 grams/liter (“gm/l”), most of them above 4.5 gm/l, and some at 7 gm/l or higher, using the fermentation conditions described in Bogosian et al., J. Biol. Chem., vol. 264, pp. 531-39, Jan. 5, 1989, except that 50 ppm of nalidixic acid was used as the inducer instead of indole acrylic acid.  
         [0012]    The expression levels of mature ST vectors are very unpredictable from published teachings, e.g., P. H. Seeburg et al. In fact, the vectors comprising the sequences of SEQ ID NOs 1 through 17, 30 through 35, and 37 through 42 have been found, using calculations as recommended in Seeburg et al., to have mRNA secondary structure greater than, rather than lessened from that of bST cDNA.  
                                     TABLE I                       SEQ ID NO   RBS   pXT                                1   dps   600       2   dps   601       3   dps   602       4   dps   658       5   dps   670       6   dps   672       7   dps   673       8   dps   674       9   dps   675       10   dps   676       11   dps   682       12   dps   683       13   dps   686       14   dps   689       15   dps   703       16   dps   709       17   dps   747       18   L49   487       19   L49   536       20   L49   541       21   L49   542       22   L49   548       23   L49   549       24   L49   562       25   L49   576       26   L49   579       27   L49   580       28   L49   581       29   L49   584       30   L437   491       31   L437   529       32   L437   530       33   L437   619       34   L437   605       35   L437   606       36   L437   607       37   L437   620       38   L437   631       39   L437   632       40   L437   633       41   L437   635       42   L437   681       43   R806   483       44   R806   564                  
 
         [0013]    [0013] 
     
       
       
         1 
         
           
             50  
           
           
             1  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 600  
             
           
            1 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagca    120 

atgagcttgt ccggactttt tgcaaatgct gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             2  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 601  
             
           
            2 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc    120 

atgtcattgt ccggactgtt tgcgaacgct gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             3  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT602  
             
           
            3 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttccctgca    120 

atgtcattgt ccggactgtt tgcaaatgcg gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             4  
             684  
             DNA  
             Artificial sequence  
             
               Somatotropin expression vector pXT 658  
             
           
            4 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagca    120 

atgtcattgt ccggactctt tgcaaatgcg gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             5  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 670  
             
           
            5 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc    120 

atgagcttgt ccggattgtt tgcaaacgct gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             6  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 672  
             
           
            6 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc    120 

atgtcattgt ccggactgtt tgcaaatgct gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             7  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 673  
             
           
            7 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc    120 

atgagcttgt ccggactgtt tgccaacgct gtcctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             8  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 674  
             
           
            8 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc    120 

atgagcttgt ccggactctt tgccaatgct gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             9  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 675  
             
           
            9 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc    120 

atgagcttgt ccggactgtt tgccaatgct gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             10  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 676  
             
           
            10 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc    120 

atgtccttgt ccggactctt tgccaacgct gtcctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             11  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 682  
             
           
            11 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc    120 

atgtcattgt ccggactgtt tgcgaacgct gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             12  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 683  
             
           
            12 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc    120 

atgtcattgt ccggactctt tgcaaatgct gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             13  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 686  
             
           
            13 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc    120 

atgtcattgt ccggactctt tgcgaacgct gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             14  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 689  
             
           
            14 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc    120 

atgtcattgt ccggactgtt tgcaaacgct gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             15  
             687  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 703  
             
           
            15 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc    120 

atgtcattgt ccggactgtt tgcgaacgct gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gcgttactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc gacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

agctggcttg ggccgctgca gttcctcagc cgtgtcttca ccaacagctt ggtgtttggc    420 

accagcgacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgtgagctgg aagatggcac cccgcgtgct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccgtaagg acctgcataa gacggagacg tacctgcgtg tcatgaagtg ccgccgcttc    660 

ggggaggcga gctgcgcctt ctaatag                                        687 

 
           
             16  
             687  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 709  
             
           
            16 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc    120 

atgagcttgt ccggactctt tgccaatgct gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gcgttactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc gacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

agctggcttg ggccgctgca gttcctcagc cgtgtcttca ccaacagctt ggtgtttggc    420 

accagcgacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgtgagctgg aagatggcac cccgcgtgct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccgtaagg acctgcataa gacggagacg tacctgcgtg tcatgaagtg ccgccgcttc    660 

ggggaggcga gctgcgcctt ctaatag                                        687 

 
           
             17  
             687  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 747  
             
           
            17 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc    120 

atgtcattgt ccggactctt tgcgaacgct gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gcgttactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc gacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

agctggcttg ggccgctgca gttcctcagc cgtgtcttca ccaacagctt ggtgtttggc    420 

accagcgacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgtgagctgg aagatggcac cccgcgtgct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccgtaagg acctgcataa gacggagacg tacctgcgtg tcatgaagtg ccgccgcttc    660 

ggggaggcga gctgcgcctt ctaatag                                        687 

 
           
             18  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 487  
             
           
            18 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttcctgcg    120 

atgtcattgt ccggattgtt tgcgaatgcg gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             19  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 536  
             
           
            19 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca    120 

atgtcattgt ccggattgtt tgcaaatgcc gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             20  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 541  
             
           
            20 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca    120 

atgtcattgt ccggattgtt tgcgaacgcc gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             21  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 542  
             
           
            21 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttcctgca    120 

atgtcattgt ccggactttt tgccaacgca gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             22  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 548  
             
           
            22 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca    120 

atgtcattgt ccggactctt tgcaaacgcc gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             23  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 549  
             
           
            23 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttcctgca    120 

atgtccttgt ccggactttt tgccaatgca gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             24  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 562  
             
           
            24 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagcc    120 

atgtcattgt ccggactttt tgccaacgca gtcctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             25  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 576  
             
           
            25 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca    120 

atgtcattgt ccggactgtt tgcgaacgcg gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             26  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 579  
             
           
            26 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca    120 

atgtcattgt ccggactttt tgccaacgcg gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             27  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 580  
             
           
            27 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca    120 

atgtcattgt ccggactgtt tgccaacgcc gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             28  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 581  
             
           
            28 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagcc    120 

atgtccttgt ccggactttt tgccaacgca gtcctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             29  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 584  
             
           
            29 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca    120 

atgtcattgt ccggactttt tgcgaatgct gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             30  
             685  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 491  
             
           
            30 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc    120 

catgtccttg tccggattgt ttgcgaatgc ggtcctccgg gctcagcacc tgcatcagct    180 

ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc    240 

catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa    300 

gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca    360 

gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg    420 

cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat    480 

gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa    540 

atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg    600 

cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt    660 

cggggaggcc agctgcgcct tctag                                          685 

 
           
             31  
             685  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 529  
             
           
            31 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc    120 

aatgagcttg tccggattgt ttgcgaatgc ggtgctccgg gctcagcacc tgcatcagct    180 

ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc    240 

catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa    300 

gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca    360 

gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg    420 

cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat    480 

gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa    540 

atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg    600 

cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt    660 

cggggaggcc agctgcgcct tctag                                          685 

 
           
             32  
             685  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 530  
             
           
            32 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttccagc    120 

catgtcattg tccggactct ttgcgaatgc ggtcctccgg gctcagcacc tgcatcagct    180 

ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc    240 

catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa    300 

gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca    360 

gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg    420 

cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat    480 

gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa    540 

atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg    600 

cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt    660 

cggggaggcc agctgcgcct tctag                                          685 

 
           
             33  
             685  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 619  
             
           
            33 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc    120 

gatgagcttg tccggattgt ttgcgaatgc agtgctccgg gctcagcacc tgcatcagct    180 

ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc    240 

catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa    300 

gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca    360 

gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg    420 

cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat    480 

gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa    540 

atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg    600 

cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt    660 

cggggaggcc agctgcgcct tctag                                          685 

 
           
             34  
             685  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 605  
             
           
            34 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttccagc    120 

aatgtccttg tccggactgt ttgcgaacgc agtcctccgg gctcagcacc tgcatcagct    180 

ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc    240 

catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa    300 

gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca    360 

gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg    420 

cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat    480 

gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa    540 

atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg    600 

cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt    660 

cggggaggcc agctgcgcct tctag                                          685 

 
           
             35  
             685  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 606  
             
           
            35 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc    120 

aatgtccttg tccggattgt ttgcgaacgc ggtcctccgg gctcagcacc tgcatcagct    180 

ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc    240 

catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa    300 

gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca    360 

gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg    420 

cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat    480 

gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa    540 

atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg    600 

cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt    660 

cggggaggcc agctgcgcct tctag                                          685 

 
           
             36  
             685  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 607  
             
           
            36 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc    120 

catgtccttg tccggactct ttgccaacgc tgtgctccgg gctcagcacc tgcatcagct    180 

ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc    240 

catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa    300 

gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca    360 

gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg    420 

cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat    480 

gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa    540 

atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg    600 

cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt    660 

cggggaggcc agctgcgcct tctag                                          685 

 
           
             37  
             685  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 620  
             
           
            37 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc    120 

aatgagcttg tccggacttt ttgcgaatgc tgtactccgg gctcagcacc tgcatcagct    180 

ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc    240 

catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa    300 

gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca    360 

gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg    420 

cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat    480 

gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa    540 

atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg    600 

cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt    660 

cggggaggcc agctgcgcct tctag                                          685 

 
           
             38  
             685  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 631  
             
           
            38 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc    120 

catgagcttg tccggattgt ttgcaaacgc cgtcctccgg gctcagcacc tgcatcagct    180 

ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc    240 

catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa    300 

gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca    360 

gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg    420 

cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat    480 

gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa    540 

atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg    600 

cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt    660 

cggggaggcc agctgcgcct tctag                                          685 

 
           
             39  
             685  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 632  
             
           
            39 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttccagc    120 

catgtcattg tccggattgt ttgcaaacgc tgtcctccgg gctcagcacc tgcatcagct    180 

ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc    240 

catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa    300 

gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca    360 

gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg    420 

cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat    480 

gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa    540 

atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg    600 

cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt    660 

cggggaggcc agctgcgcct tctag                                          685 

 
           
             40  
             685  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 633  
             
           
            40 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttccagc    120 

catgtcattg tccggacttt ttgcgaacgc tgtgctccgg gctcagcacc tgcatcagct    180 

ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc    240 

catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa    300 

gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca    360 

gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg    420 

cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat    480 

gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa    540 

atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg    600 

cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt    660 

cggggaggcc agctgcgcct tctag                                          685 

 
           
             41  
             685  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 635  
             
           
            41 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc    120 

catgtcattg tccggacttt ttgccaatgc ggtcctccgg gctcagcacc tgcatcagct    180 

ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc    240 

catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa    300 

gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca    360 

gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg    420 

cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat    480 

gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa    540 

atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg    600 

cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt    660 

cggggaggcc agctgcgcct tctag                                          685 

 
           
             42  
             685  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 681  
             
           
            42 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgttcccagc    120 

catgtcattg tccggacttt ttgccaacgc ggtgctccgg gctcagcacc tgcatcagct    180 

ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc    240 

catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa    300 

gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca    360 

gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg    420 

cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat    480 

gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa    540 

atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg    600 

cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt    660 

cggggaggcc agctgcgcct tctag                                          685 

 
           
             43  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 483  
             
           
            43 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcctaaaaga gactaggagg agattagaat gttcccagca    120 

atgagtttgt ccggactgtt tgcgaacgcc gtgctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             44  
             684  
             DNA  
             Artificial Sequence  
             
               Somatotropin expression vector pXT 564  
             
           
            44 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcctaaaaga gactaggagg agattagaat gttcccagca    120 

atgtcattgt ccggactgtt tgccaatgcg gtcctccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctag                                           684 

 
           
             45  
             25  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            45 

cataacatca agaggatatg aaatt                                           25 

 
           
             46  
             25  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            46 

gtacaaatca tagagggtat ttaat                                           25 

 
           
             47  
             26  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            47 

agacactaaa tagagggtat ttaatt                                          26 

 
           
             48  
             25  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            48 

taaaagagac taggaggaga ttaga                                           25 

 
           
             49  
             983  
             DNA  
             Artificial Sequence  
             
               Synthetic construct for expression of bovine 
      somatotropin  
             
           
            49 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc    120 

atgagcttgt ccggactctt tgccaatgct gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc    420 

acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc    660 

ggggaggcca gctgcgcctt ctagaagctt aattctcact cattaggcac cccaggcttt    720 

acactttatg cttccggctc gtataatgtg tggaattgtg agcggataac aatttcacac    780 

aggaaacagc tgattgccct tcaccgcctg gcctccgttg agccatctgg atcggcagcg    840 

ttgtcttcat caaccggaac gagcatgccg gagagcagct cactcattag gcaccccagg    900 

ctttacactt tatgcttccg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc    960 

acacaggaaa cagaattaag ctt                                            983 

 
           
             50  
             977  
             DNA  
             Artificial Sequence  
             
               Synthetic construct for expression of bovine 
      somatotropin  
             
           
            50 

gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat     60 

ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc    120 

atgagcttgt ccggactctt tgccaatgct gtactccggg ctcagcacct gcatcagctg    180 

gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gcgttactcc    240 

atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc gacgggcaag    300 

aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag    360 

agctggcttg ggccgctgca gttcctcagc cgtgtcttca ccaacagctt ggtgtttggc    420 

accagcgacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg    480 

cgtgagctgg aagatggcac cccgcgtgct gggcagatcc tcaagcagac ctatgacaaa    540 

tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc    600 

ttccgtaagg acctgcataa gacggagacg tacctgcgtg tcatgaagtg ccgccgcttc    660 

ggggaggcga gctgcgcctt ctaatagctc gagtctagac tcactcatta ggcaccccag    720 

gctttacact ttatgcttcc ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt    780 

cacacaggaa aaagctgatt gcccttcacc gcctggcctc cgttgagcca tctggatcgg    840 

cagcgttgtc ttcatcaacc ggaacgagca tgccggagag cagctcactc attaggcacc    900 

ccaggcttta cactttatgc ttccggctcg tataatgtgt ggaattgtga gcggataaca    960 

atttcacaca gaagctt                                                   977