DNA SEQUENCES FOR AN AMINO ACID TRANSPORTER, PLASMIDS, BACTERIA, YEASTS AND PLANTS CONTAINING A TRANSPORTER AND THEIR USE

Abstract: There are described DNA sequences, that contain the coding region of amino acid transporters, whose introduction in a plant genome modifies the transfer of metabolites in transgenic plants, plasmids, bacteria, yeasts and plants containing these DNA sequences, as well as their use.

Description: Title: DNA sequences for an amino acid transporter- plasmids, bacteria, yeasts and plants containing a transporter and their use Field of the invention The present invention relates to DNA sequences, that contain the coding region of amino acid transporters, whose introduction in a plant genome modifies the transfer of metabolites in transgenic plants, plasmids, bacteria, yeasts and plants containing these DNA sequences, as well as their use. For many plant species it is known, that the delivery of energy rich compounds to the phloem through the cell wall takes place throughout the cell. Transporter molecules, which allow the penetration of amino acids through the plant cell wall are not known. In bacteria, numerous amino acid transport systems have been characterised. For aromatic amino acids, 5 different transporters have been described, which can transport one of phenylalanine, tyrosine and tryptophan, whilst the others are specific for individual amino acids, (see Sarsero et al., 1991, J Bacteriol 173: 3231-3234). The speed constants of the transport process indicates that the specific transport is less efficient. For several transporter proteins, the corresponding genes have been cloned. This has been achieved using transport deficient mutants, which were selected for their transport ability after transformation with DNA fragments as inserts in expression vectors, (see Wallace et al., 1990, J Bacteriol 172: 3214-3220). The mutants were selected depending on their ability to grow in the presence of toxic analogues of amino acids, since they cannot take up these and therefore cannot be impaired. Corresponding complementation studies have been carried out with the eukaryotic yeast, Saccharomyces cereviεiae . Tanaka & Fink (1985, Gene 38: 205-214) describe a histidine transporter that was identified by complementation of a mutation. Vandenbol et al. (1989, Gene 83: 153-159) describe a proline transporter for Saccharomyces cereviεiae . The yeast possesses two different permeases for proline. One transports with lower efficiency and can be used also for other amino acids, and the other is proline-specific and works with high affinity. The latter was coded from the put4 gene. This carries an open reading frame for a peptide with a molecular weight of 69 kDa. The protein contains 12 membrane-penetrating regions, but does not contain any N- terminal signal sequence for secretion. This is a typical property of integral membrane proteins. The permeases possess homology for arginine and for histidine permease from yeast, but not however for proline permease from Escherichia coli . For plant cells, by studies on tobacco suspension cultures, it has been found that the transport of arginine, asparagine, phenylalanine and histidine are pH and energy dependent. Since a 1000-fold excess of leucine inhibits the transport of the other amino acids, it can be assumed therefore that all use the same transporter (McDaniel et al., 1982., Plant Physio 69: 246-249). Li and Bush (1991, Plant Physiol 96: 1338-1344) determined for aliphatic, neutral amino acids two transport systems in plasma membrane vesicles from Beta vulgaris . On the one hand, alanine, methionine, glutamine and leucine displace each other on the transporter protein, and on the other hand, isoleucine, valine and threonine have mutually competitive effects. In combined competition kinetic studies (Li & Bush, 1990, Plant Physiol 94: 268-277) four different transport systems have been distinguished. Besides a transporter for all neutral amino acids, which work with low affinity, there exists a high affinity type which, however, possesses low affinity to isoleucine, threonine, valine and proline. Further transporters exist for acids as well as for basic amino acids. The transporter molecule or gene for plant transporter proteins is not known. There are now described DNA sequences which contain the coding region of a plant amino acid transporter, and whose information contained in the nucleotide sequence allows, by integration in a plant genome, the formation of RNA, by which a new amino acid transport activity can be introduced in the plant cells or an endogenous amino acid transporter activity can be expressed. Under the term amino transporter is to be understood, for example a cDNA sequence that codes an amino transporter from Arabidopsis thaliana . The identification of the coding region of the amino acid transporter is carried out by a process which allows the isolation of plant DNA sequences which code transporter molecules by means of expression in specific mutants of yeast Saccharomyces cerevisiae . For this, suitable yeast mutants have to be provided which cannot take up a substance for which the coding region of a transporter molecule has to be isolated from a plant gene library. A mutant which cannot grow in media, with proline or citrulline as the only nitrogen source, is described by Jauniaux et al. (1987), Eur J Biochem 164: 601-606). For the preparation of yeast strains that can be used to 5 identify plant amino acid transporters, a yeast mutant which is not able to grow in media with proline and/or citrulline as the only nitrogen source, is for example transformed with pFL 61 plasmid, which carries, as an insert, cDNA fragments from a cDNA library from 10 Arabidopsis thaliana . Further, a double mutant JT16 (Tanaka & Fink, 1985, Gene 38: 205-214) which has a deficiency in histidine synthesis (his4 ) and in histidine uptake (hipl ) is transformed with 15 the described pFL 61 plasmid and cultivated in a medium with addition of histidine. It has now surprisingly been found that, in the transformation of yeast cells, certain plant cDNA 20 fragments can complement the yeast mutation. By analysis of the properties of the proteins coded from the cDNA it can be shown that for the complementing of the mutation, a coding region is responsible that codes a plant amino acid transporter with a wide specificity spectrum, (see example 25 3) . Such a coding region of an amino acid transporter is shown foe example by one of the following nucleotide sequences: 1. Sequence (Seq. ID No. 1) : CTTAAAACAT TTATTTTATC TTCTTCTTGT TCTCTCTTTC TCTTTCTCTC ATCACT 56 ATG AAG AGT TTC AAC ACA GAA GGA CAC AAC CAC TCC ACG GCG GAA 101 Met Lys Ser Phe Asn Thr Glu Gly His Asn His Ser Thr Ala Glu 1 5 10 15 TCC GGC GAT GCC TAC ACC GTG TCG GAC CCG ACA AAG AAC GTC GAT 146 Ser Gly Asp Ala Tyr Thr Val Ser Asp Pro Thr Lys Asn Val Asp 20 25 30 GAA GAT GGT CGA GAG AAG CGT ACC GGG ACG TGG CTT ACG GCG AGT 191 Glu Asp Gly Arg Glu Lys Arg Thr Gly Thr Trp Leu Thr Ala Ser 35 40 45 GCG CAT ATT ATC ACG GCG GTG ATA GGC TCC GGA GTG TTG TCT TTA 236 Ala His lie lie Thr Ala Val lie Gly Ser Gly Val Leu Ser Leu 50 55 60 GCA TGG GCT ATA GCT CAG CTT GGT TGG ATC GCA GGG ACA TCG ATC 281 Ala Trp Ala lie Ala Gin Leu Gly Trp lie Ala Gly Thr Ser lie 65 70 75 TTA CTC ATT TTC TCG TTC ATT ACT TAC TTC ACC TCC ACC ATG CTT 326 Leu Leu lie Phe Ser Phe lie Thr Tyr Phe Thr Ser Thr Met Leu 80 85 90 GCC GAT TGC TAC CGT GCG CCG GAT CCC GTC ACC GGA AAA CGG AAT 371 Ala Asp Cys Tyr Arg Ala Pro Asp Pro Val Thr Gly Lys Arg Asn 95 100 105 TAC ACT TAC ATG GAC GTT GTT CGA TCT TAC CTC GGT GGT AGG AAA 416 Tyr Thr Tyr Met Asp Val Val Arg Ser Tyr Leu Gly Gly Arg Lys 110 115 120 GTG CAG CTC TGT GGA GTG GCA CAA TAT GGG AAT CTG ATT GGG GTC 461 Val Gin Leu Cys Gly Val Ala Gin Tyr Gly Asn Leu lie Gly Val 125 130 135 ACT GTT GGT TAC ACC ATC ACT GCT TCT ATT AGT TTG GTA GCG GTA 506 Thr Val Gly Tyr Thr lie Thr Ala Ser lie Ser Leu Val Ala Val 140 145 150 GGG AAA TCG AAC TGC TTC CAC GAT AAA GGG CAC ACT GCG GAT TGT 551 Gly Lys Ser Asn Cys Phe His Asp Lys Gly His Thr Ala Asp Cys 155 160 165 ACT ATA TCG AAT TAT CCG TAT ATG GCG GTT TTT GGT ATC ATT CAA 596 Thr lie Ser Asn Tyr Pro Tyr Met Ala Val Phe Gly lie lie Gin 170 175 180 GTT ATT CTT AGC CAG ATC CCA AAT TTC CAC AAG CTC TCT TTT CTT 641 Val lie Leu Ser Gin lie Pro Asn Phe His Lys Leu Ser Phe Leu 185 190 195 TCC ATT ATG GCC GCA GTC ATG TCC TTT ACT TAT GCA ACT ATT GGA 686 Ser lie Met Ala Ala Val Met Ser Phe Thr Tyr Ala Thr lie Gly 200 205 210 ATC GGT CTA GCC ATC GCA ACC GTC GCA GGT GGG AAA GTG GGT AAG 731 lie Gly Leu Ala lie Ala Thr Val Ala Gly Gly Lys Val Gly Lys 215 220 225 ACG AGT ATG ACG GGC ACA GCG GTT GGA GTA GAT GTA ACC GCA GCT 776 Thr Ser Met Thr Gly Thr Ala Val Gly Val Asp Val Thr Ala Ala 230 235 240 CAA AAG ATA TGG AGA TCG TTT CAA GCG GTT GGG GAC ATA GCG TTC 821 Gin Lys lie Trp Arg Ser Phe Gin Ala Val Gly Asp lie Ala Phe 245 250 255 GCC TAT GCT TAT GCC ACG GTT CTC ATC GAG ATT CAG GAT ACA CTA 866 Ala Tyr Ala Tyr Ala Thr Val Leu lie Glu lie Gin Asp Thr Leu 260 265 270 AGA TCT AGC CCA GCT GAG AAC AAA GCC ATG AAA AGA GCA AGT CTT 911 Arg Ser Ser Pro Ala Glu Asn Lys Ala Met Lys Arg Ala Ser Leu 275 280 285 GTG GGA GTA TCA ACC ACC ACT TTT TTC TAC ATC TTA TGT GGA TGC 956 Val Gly Val Ser Thr Thr Thr Phe Phe Tyr lie Leu Cys Gly Cyε 290 295 300 ATC GGC TAT GCT GCA TTT GGA AAC AAT GCC CCT GGA GAT TTC CTC 1001 lie Gly Tyr Ala Ala Phe Gly Asn Asn Ala Pro Gly Asp Phe Leu 305 310 315 ACA GAT TTC GGG TTT TTC GAG CCC TTT TGG CTC ATT GAC TTT GCA 1046 Thr Asp Phe Gly Phe Phe Glu Pro Phe Trp Leu lie Asp Phe Ala 320 325 330 AAC GCT TGC ATC GCT GTC CAC CTT ATT GGT GCC TAT CAG GTG TTC 1091 Asn Ala Cys lie Ala Val His Leu lie Gly Ala Tyr Gin Val Phe 335 340 345 GCG CAG CCG ATA TTC CAG TTT GTT GAG AAA AAA TGC AAC AGA AAC 1136 Ala Gin Pro lie Phe Gin Phe Val Glu Lys Lys Cys Asn Arg Asn 350 355 360 TAT CCA GAC AAC AAG TTC ATC ACT TCT GAA TAT TCA GTA AAC GTA 1181 Tyr Pro Asp Asn Lys Phe lie Thr Ser Glu Tyr Ser Val Asn Val 365 370 375 CCT TTC CTT GGA AAA TTC AAC ATT AGC CTC TTC AGA TTG GTG TGG 1226 Pro Phe Leu Gly Lys Phe Asn lie Ser Leu Phe Arg Leu Val Trp 380 385 390 AGG ACA GCT TAT GTG GTT ATA ACC ACT GTT GTA GCT ATG ATA TTC 1271 Arg Thr Ala Tyr Val Val lie Thr Thr Val Val Ala Met lie Phe 395 400 405 CCT TTC TTC AAC GCG ATC TTA GGT CTT ATC GGA GCA GCT TCC TTC 1316 Pro Phe Phe Asn Ala lie Leu Gly Leu lie Gly Ala Ala Ser Phe 410 415 420 δ TGG CCT TTA ACG GTT TAT TTC CCT GTG GAG ATG CAC ATT GCA CAA 1361 Trp Pro Leu Thr Val Tyr Phe Pro Val Glu Met His He Ala Gin 425 430 435 ACC AAG ATT AAG AAG TAC TCT GCT AGA TGG ATT GCG CTG AAA ACG 1406 Thr Lys He Lys Lys Tyr Ser Ala Arg Trp He Ala Leu Lys Thr 440 445 450 ATG TGC TAT GTT TGC TTG ATC GTC TCG CTC TTA GCT GCA GCC GGA 1451 Met Cys Tyr Val Cys Leu He Val Ser Leu Leu Ala Ala Ala Gly 455 460 465 TCC ATC GCA GGA CTT ATA AGT AGT GTC AAA ACC TAC AAG CCC TTC 1496 Ser He Ala Gly Leu He Ser Ser Val Lys Thr Tyr Lys Pro Phe 470 475 480 CGG ACT ATG CAT GAG TGAGTTTGAG ATCCTCAAGA GAGTCAAAAA 1541 Arg Thr Met His Glu 485 TATATGTAGT AGTTTGGTCT TTCTGTTAAA CTATCTGGTG TCTAAATCCA 1591 ATGAGAATGC TTTATTGCTA AAACTTCATG AATCTCTCTG TATCTACATC 1641 TTTCAATCTA ATACATATGA GCTCTTCCAA AAAAAAAAAA AAAA 1685 2. Sequence (Seq. ID No. 2): CTATTTTAT AATTCCTCTT CTTTTTGTTC 29 ATAGCTTTGT AATTATAGTC TTATTTCTCT TTAAGGCTCA ATAAGAGGAG 79 ATG GGT GAA ACC GCT GCC GCC AAT AAC CAC CGT CAC CAC CAC CAT 124 Met Gly Glu Thr Ala Ala Ala Asn Asn His Arg His His His His 1 5 10 15 CAC GGC CAC CAG GTC TTT GAC GTG GCC AGC CAC GAT TTC GTC CCT 169 His Gly His Gin Val Phe Asp Val Ala Ser His Asp Phe Val Pro 20 25 30 CCA CAA CCG GCT TTT AAA TGC TTC GAT GAT GAT GGC CGC CTC AAA 214 Pro Gin Pro Ala Phe Lys Cys Phe Asp Asp Asp Gly Arg Leu Lys 35 40 45 AGA ACT GGG ACT GTT TGG ACC GCG AGC GCT CAT ATA ATA ACT GCG 259 Arg Thr Gly Thr Val Trp Thr Ala Ser Ala His He He Thr Ala 50 55 60 GTT ATC GGA TCC GGC GTT TTG TCA TTG GCG TGG GCG ATT GCA CAG 304 Val He Gly Ser Gly Val Leu Ser Leu Ala Trp Ala He Ala Gin 65 70 75 CTC GGA TGG ATC GCT GGC CCT GCT GTG ATG CTA TTG TTC TCT CTT 349 Leu Gly Trp He Ala Gly Pro Ala Val Met Leu Leu Phe Ser Leu 80 85 90 GTT ACT CTT TAC TCC TCC ACA CTT CTT AGC GAC TGC TAC AGA ACC 394 Val Thr Leu Tyr Ser Ser Thr Leu Leu Ser Asp Cys Tyr Arg Thr 95 100 105 GGC GAT GCA GTG TCT GGC AAG AGA AAC TAC ACT TAC ATG GAT GCC 439 Gly Asp Ala Val Ser Gly Lys Arg Asn Tyr Thr Tyr Met Asp Ala 110 115 120 GTT CGA TCA ATT CTC GGT GGG TTC AAG TTC AAG ATT TGT GGG TTG 484 Val Arg Ser He Leu Gly Gly Phe Lys Phe Lys He Cys Gly Leu 125 130 135 ATT CAA TAC TTG AAT CTC TTT GGT ATC GCA ATT GGA TAC ACG ATA 529 He Gin Tyr Leu Asn Leu Phe Gly He Ala He Gly Tyr Thr He 140 145 150 GCA GCT TCC ATA AGC ATG ATG GCG ATC AAG AGA TCC AAC TGC TTC 574 Ala Ala Ser He Ser Met Met Ala He Lys Arg Ser Asn Cys Phe 155 160 165 CAC AAG AGT GGA GGA AAA GAC CCA TGT CAC ATG TCC AGT AAT CCT 619 His Lys Ser Gly Gly Lys Asp Pro Cys His Met Ser Ser Asn Pro 170 175 180 TAC ATG ATC GTA TTT GGT GTG GCA GAG ATC TTG CTC TCT CAG GTT 664 Tyr Met He Val Phe Gly Val Ala Glu He Leu Leu Ser Gin Val 185 190 200 CCT GAT TTC GAT CAG ATT TGG TGG ATC TCC ATT GTT GCA GCT GTT 709 Pro Asp Phe Asp Gin He Trp Trp He Ser He Val Ala Ala Val 205 210 220 ATG TCC TTC ACT TAC TCT GCC ATT GGT CTA GCT CTT GGA ATC GTT 754 Met Ser Phe Thr Tyr Ser Ala He Gly Leu Ala Leu Gly He Val 225 230 235 CAA GTT GCA GCG AAT GGA GTT TTC AAA GGA AGT CTC ACT GGA ATA 799 Gin Val Ala Ala Asn Gly Val Phe Lys Gly Ser Leu Thr Gly He 240 245 250 AGC ATC GGA ACA GTG ACT CAA ACA CAG AAG ATA TGG AGA ACC TTC 844 Ser He Gly Thr Val Thr Gin Thr Gin Lys He Trp Arg Thr Phe 255 260 265 CAA GCA CTT GGA GAC ATT GCC TTT GCG TAC TCA TAC TCT GTT GTC 889 Gin Ala Leu Gly Asp He Ala Phe Ala Tyr Ser Tyr Ser Val Val 270 275 280 CTA ATC GAG ATT CAG GAT ACT GTA AGA TCC CCA CCG GCG GAA TCG 934 Leu He Glu He Gin Asp Thr Val Arg Ser Pro Pro Ala Glu Ser 285 290 295 AAA ACG ATG AAG AAA GCA ACA AAA ATC AGT ATT GCC GTC ACA ACT 979 Lys Thr Met Lys Lys Ala Thr Lys He Ser He Ala Val Thr Thr 300 305 310 ATC TTC TAC ATG CTA TGT GGC TCA ATG GGT TAT GCC GCT TTT GGA 1024 He Phe Tyr Met Leu Cys Gly Ser Met Gly Tyr Ala Ala Phe Gly 315 320 325 GAT GCA GCA CCG GGA AAC CTC CTC ACC GGT TTT GGA TTC TAC AAC 1069 Asp Ala Ala Pro Gly Asn Leu Leu Thr Gly Phe Gly Phe Tyr Asn 330 335 340 CCG TTT TGG CTC CTT GAC ATA GCT AAC GCC GCC ATT GTT GTC CAC 1114 Pro Phe Trp Leu Leu Asp He Ala Asn Ala Ala He Val Val His 245 350 355 CTC GTT GGA GCT TAC CAA GTC TTT GCT CAG CCC ATC TTT GCC TTT 1159 Leu Val Gly Ala Tyr Gin Val Phe Ala Gin Pro He Phe Ala Phe 360 365 370 ATT GAA AAA TCA GTC GCA GAG AGA TAT CCA GAC AAT GAC TTC CTC 1204 He Glu Lys Ser Val Ala Glu Arg Tyr Pro Asp Asn Asp Phe Leu 375 380 385 AGC AAG GAA TTT GAA ATC AGA ATC CCC GGA TTT AAG TCT CCT TAC 1249 Ser Lys Glu Phe Glu He Arg He Pro Gly Phe Lys Ser Pro Tyr 390 395 400 AAA GTA AAC GTT TTC AGG ATG GTT TAC AGG AGT GGC TTT GTC GTT 1294 Lys Val Asn Val Phe Arg Met Val Tyr Arg Ser Gly Phe Val Val 405 410 415 ACA ACC ACC GTG ATA TCG ATG CTG ATG CCG TTT TTT AAC GAC GTG 1339 Thr Thr Thr Val He Ser Met Leu Met Pro Phe Phe Asn Asp Val 420 425 430 GTC GGG ATC TTA GGG GCG TTA GGG TTT TGG CCC TTG ACG GTT TAT 1384 Val Gly He Leu Gly Ala Leu Gly Phe Trp Pro Leu Thr Val Tyr 435 440 445 TTT CCG GTG GAG ATG TAT ATT AAG CAG AGG AAG GTT GAG AAA TGG 1429 Phe Pro Val Glu Met Tyr He Lys Gin Arg Lys Val Glu Lys Trp 450 455 460 AGC ACG AGA TGG GTG TGT TTA CAG ATG CTT AGT GTT GCT TGT CTT 1474 Ser Thr Arg Trp Val Cys Leu Gin Met Leu Ser Val Ala Cys Leu 465 470 475 GTG ATC TCG GTG GTC GCC GGG GTT GGA TCA ATC GCC GGA GTG ATG 1519 Val He Ser Val Val Ala Gly Val Gly Ser He Ala Gly Val Met 480 485 490 CTT GAT CTT AAG GTC TAT AAG CCA TTC AAG TCT ACA TAT 1558 Leu Asp Leu Lys Val Tyr Lys Pro Phe Lys Ser Thr Tyr 495 500 TGATGATTAT GGACCATGAA CAACAGAGAG AGTTGGTGTG TAAAGTTTAC 1608 CATTTCAAAG AAAACTCCAA AAATGTGTAT ATTGTATGTT GTTCTCATTT 1658 CGTATGGTCT CATCTTTGTA ATAAAATTTA AAACTTATGT TATAAATTAT 1708 AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AA 1740 The DNA sequences of the invention identified with the help of the transformed yeast strains, e.g sequences Seq. No. 1 and No. 2, can be introduced into plasmids and thereby be combined with steering elements for expression in eukaryotic cells (see Example 4) . These steering elements are on the one handed transcription promoters, and on the other hand transcription terminators. With the plasmids, eukaryotic cells can be transformed, with the aim of expression of a translatable mRNA which makes possible the synthesis of an amino acid transporter in the cells or with the aim of expression of a non-translatable RNA, which prevents synthesis of an endogenous amino acid transporter in the cells. By expression of an RNA corresponding to the inventive sequences of plant amino acid transporters, a modification of the plant acid metabolism, as well as total nitrogen metabolism, is possible, the economic significance of which is obvious. Nitrogen is the nutrient mainly responsible for limiting growth. The viability of germ lines as well as germination capacity of seeds is directly dependent on the nitrogen content of storage tissue. The formation of high value food materials with a high protein content is dependent on a sufficient nitrogen supply. Nitrogen is transported essentially in the form of amino acids. An improvement in the delivery of amino acids to their harvested parts can therefore lead to an increase in yield of agricultural plants. The possibility of forcing the take-up of ar-ino acid in individual organs allows the qualitative improvement of such organs, which because of the demands of the utilization process, contain little nitrogen, for example potatoes which are grown for the production of starch. Besides this, modifications of the whole plant by which the growth of individual tissues, for example leaves, is slowed down, whilst the growth of the harvested parts is increased. For this, one can imagine a lengthening of the vegetative phase of crops, which leads to an increased formation of storage substances. Processes for the genetic modification of dicotyledonous and onocotyledonous plants are already known, (see for example Gasser, C.S., Fraley, R.T., 1989, Science 244: 1293-1299; Potrykus, 1991, Ann Rev Plant Mol Biol Plant Physiol 42: 205-225) . For expression in plants - the coding sequences must be coupled with the transcriptional regulatory elements. Such elements called promoters, are known (EP 375091) . Further, the coding regions must be provided with transcription termination signals with which they can be correctly transcribed. Such elements are also described (see Gielen et al., 1989, EMBO J 8: 23-29). The transcriptional start region can be both native and/or homologous as well as foreign and/or heterologous to the host plant. If desired, termination regions are interchangeable with one another. The DNA sequence of the transcription starting and termination regions can be prepared synthetically or obtained naturally, or obtained from a mixture of synthetic and natural DNA constituents. For introduction of foreign genes in higher plants a large number of cloning vectors are available that include a replication signal for E. coli and a marker which allows a selection of the transformed cells. Examples of such vectors are pBR 322, pUC-Series, M13 mp-Series, pACYC 184 etc. Depending on the method of introduction of the desired gene in the plants, other DNA sequences may be suitable. Should the Ti- or Ri-plasmid be used, e.g. for the transformation of the plant cell, then at least the right boundary, often however both the right and left boundary of the Ti- and Ri-Plasmid T-DNA, is attached, as a flanking region, to the gene being introduced. The use of T-DNA for the transformation of plants cells has been intensively researched and is well described in EP 120 516; Hoeka a, In: The Binary Plant Vector System, Offset-drukkerij Kanters B.V. Alblasserdam, (1985) , Chapter V; Fraley, et al., Crit. Rev. Plant Sci., 4:1-46 and An et al. (1985) EMBO J. 4: 277-287. Once the introduced DNA is integrated in the genome, it is as a rule stable there and remains also in the offspring of the original transformed cells. It normally contains a selection marker, which induces resistance in the transformed plant cells against a biocide or antibiotic such as kanamycin, G 418, bleomycin, hygromycin or phosphinotricin etc. The individual marker employed should therefore allow the selection of transformed cells from cells, which lack the introduced DNA. For the introduction of DNA into a plant host cell, besides transformation using Agrohacteria, there are many other techniques available. These techniques include the fusion of protoplasts, microinjection of DNA and electroporation, as well as ballistic methods and virus infection. From the transformed plant material, whole plants can be regenerated in a suitable medium, which contains antibiotics or biocides for the selection. The resulting plants can then be tested for the presence of introduced DNA. No special demands are placed on the plasmids in injection and electroporation. Simple plasmids, such as e.g. pUC-derivatives can be used. Should however whole plants be regenerated from such transformed cells the presence of a selectable marker gene is necessary. The transformed cells grow within the plants in the usual manner (see also McCormick et al. (1986) Plant Cell Reports 5: 81-84) . These plants can be grown normally and crossed with plants, that possess the same transformed genes or different. The resulting hybrid individuals have the corresponding phenotypical properties. The DNA sequences of the invention can also be introduced in plasmids and thereby combined with steering elements for an expression in prokaryotic cells. The formation of a translatable RNA sequence of a eukaryotic amino acid transporter from bacteria leads, in spite of the considerable differences in the membrane structures of prokaryotes and eukaryotes, means in addition surprisingly, that prokaryotes can now use a eukaryotic amino acid transporter with specificity for certain substrates. This makes possible the production of bacterial strains, which could be used for studies of the properties of the transporter as well as its substrate. The invention also relates to bacteria, that contain the plasmids of the invention. The DNA sequences of the invention can also be introduced in plasmids which allow mutagenesis or a sequence modification through recombination of DNA sequences in prokaryotic or eukaryotic systems. In this way the specificity of the amino acid transporter can be modified. Thus the specificity of the transporter can be changed. The invention also relates to derivatives or parts of plasmids, that contain the DNA sequences of the invention and which can be used for the transformation of prokaryotic and eukaryotic cells. By using standard processes (see Sambrook et al., 1989, Molecular Cloning: A Laboratory Manual, 2. Edn. , Cold Spring Harbor Laboratory Press, NY, USA) . Base exchanges can be carried out or natural or synthetic sequences can be added. For binding DNA fragments with one another adaptors or linkers can be introduced on the fragments. Further, manipulations can be carried which prepare suitable restriction cleavage sides or remove the excess DNA or restriction cleavage sites. Where insertions, deletions or substitutions such as for example transition-:- and transversions are to be carried, in vitro mutagenesis, primer repair, restrictions or ligations can be used. For methods of analysis, in general a sequence analysis, restriction analysis and other biochemical molecular biological methods can be used. After each manipulation, the DNA sequence, used, can be cleaved and bound with another DNA sequence. Each plasmid sequence can be cloned in the same or different plasmids. Derivatives or parts of the DNA sequences and plasmids of the invention can also be used for the transformation of prokaryotic and eukaryotic cells. Further, the DNA sequences of the invention can be used according to standard processes for the isolation of similar sequences on the genome of plants of various species, which also code for amino acid or other oligosaccharide transporter molecules. With these sequences, constructs for the transformation of plant cells can be prepared which modify the transport process in transgenic plants. In order to specify related DNA sequences, gene libraries must first be prepared, which are representative for the content in genes of a plant type or for the expression of genes in a plant type. The former are genomic libraries, whilst the latter are cDNA libraries. From these, related sequences can be isolated using the DNA sequences of the invention as probes. Once the related gene has been identified and isolated, a determination of the sequence and an analysis of the properties of the proteins coded from this sequence is possible. In order to understand the examples forming the basis of this invention all the processes necessary for these tests and which are known per se will first of all be listed: 1. Clonincf process For cloning in E. coli , the vector pBluescriptSK (Short et al., 1988, Nucl Acids Res 16: 7583-7600) was used. For the transformation of yeasts, the vector pFL61 (Minet & Lacroute, 1990, Curr Genet 18: 287-291) was used. For the plant transformation the gene constructs in the binary vector pBIN-Hyg were cloned. 2. Bacterial and yeast strains For the pBluescriptSK vector as well as for PBinAR constructs, the E. coli strain XLlblue (Bullock et al., 1987, Biotechniques, 5, 376- 378) was used. As starting strain for the expression of the cDNA library in yeast, the yeast strain 22574d (Jauniaux et al., 1987, Eur J Biochem 164: 601-606) was used. The transformation of the plasmids in potato plants was carried out using -Agrro.bacteriu-7. tumefaciens strain LBA4404 (Bevan (1984) Nucl. Acids Res 12: 8711-8720). 3. Transformation of AσroJbacteriuπ. tumefaciens The transfer of the DNA in the Agrohacteria was carried out by direct transformation by the method of Hδfgen & Willmitzer (1988, Nucleic Acids Res 16: 9877) . The plasmid DNA of the transformed Agrobacterium was isolated in accordance with the method of Birnboim and Doly (1979) (Nucl Acids Res 7: 1513-1523) and was analysed by gel electrophoresis after suitable restriction cleavage. 4. Plant transformation Ten small leaves, wounded with a scalpel, of a sterile potato culture were placed in 10 ml of MS medium with 2% amino acid containing 30-50 μl of an Agrobacterium tumefaciens overnight culture grown under selection. After 3-5 minutes gentle shaking, the leaves were laid out on MS medium of 1.6% glucose, 2 mg/1 of zeatin ribose, 0.02 mg/1 of naphthylacetic acid, 0.02 mg/1 of gibberellic acid, 500 mg/1 of claforan, 50 mg/1 of kana ycin and 0.8% bacto agar. After incubation for one week at 25°C and 3000 lux, the claforan concentration in the medium was reduced by half. __ Deposits The following plasmids and yeast strains were deposited at the Deutschen Sammlung von Mikroorganismen (DSM) in Braunschweig, Germany on the 12.06.1992 (deposit number): Plasmid pPPPl-20 (DSM 7129) Plasmid pBinPPPl-20 (DSM 7130) Description of the Figures Fig. 1 shows the plasmid pPPPl-20, that contains the sequence Seq-ID No 1. The finely drawn line corresponds to the sequence from pBluescriptSK. The thicker line represents the cDNA insert. The cleavage positions of the inserts are shown. Fig. 2 shows the uptake of 14 C-proline from the medium. no = time perio of the uptake without competitor proline = time period with fourfold excess of unlabelled proline citrulline = time period with fourfold excess of unlabelled citrulline GABA = time period with fourfold excess of gamma-aminobutyric acid time = time in seconds cpm = decays counted per minute Fig. 3 shows the plasmid pAAP2, that contains the sequence Seq-ID No 2. The finely drawn line corresponds to the sequence from pBluescriptSK. The thicker line represents the cDNA insert. The cleavage positions of the inserts are shown. Fig. 4 shows a competition experiment with the yeast line 22574d: :AAP2, in which the uptake of 14 C labelled L-proline from the medium in the presence of a fourfold excess of other amino acids or their analogues is measured. Besides the standard abbreviations for amino acids in the three letter code the following are also used: Cit = citrulline; D-Pro = D-proline; OH-Pro = hydroxyproline and A2C = azetidine-2-carboxylic acid. Fig. 4 shows a competition experiment with the yeast line JT16: :AAP2, in which the uptake of 14 C labelled L-histidine from the medium in the presence of a tenfold excess of other amino acids or their analogues is measured. Besides the standard abbreviations for amino acids in the three letter code the following are also used: Cit = citrulline; Orn = ornithine; Can = canavanine; and NH4 = ammonium The following examples describe the cloning and identification as well as the function and use of a plant amino acid transporter. Example 1 Cloning of the cDNA of a plant amino acid transporter For complementation of the proline transport mutation of the yeast strain 22574d (Jauniaux et al., 1987, Eur J Biochem 164: 601-606) and/or the histidine synthesis and transport mutation of the strain JT16 (Tanaka & Fink, 1985, Gene 38: 205-214), there was used a cDNA of young germ lines from Arabidopsis thaliana (two leaf stage) in the yeast expression vector pFL61 (Minet & Lacroute, 1990, Curr Genet 18: 287-291) which had been made available by Minet (Minet et al. , 1992, Plant J 2: 417-422). Around 1 μg of the vector with the cDNA-insert was transformed in the yeast strain 22574d and/or JT16 by the method of Dohmen et al. (1991, Yeast 7: 691-692). Yeast transformands, which could grow in media with 4 mM proline as the sole nitrogen source or in media with 6 mM histidine, were propagated. From the lines plasmid-DNA was prepared by standard methods. Clones that could complement the particular mutation, contained plasmids with similar restriction type of the cDNA insert. These varied in size between 1.6 and 1.7 kb. Example 2 Sequence analyses of the cDNA insert of the plasmid pFL61-pppl-20 From a yeast line PPP1-20, obtained in a similar manner to example 1, which, in spite of the 22574d mutation could grow with proline as the only nitrogen source, the plasmid pFL61-pppl-20 was isolated and its cDNA insert prepared as a Notl fragment and cloned in the vector pBluescriptSK. In this way, the plasmid pPPPl-20 was obtained (see Figure 1) . Using synthetic oligonucleotides, the insert was sequenced by the method of Sanger et al. (1977, Proc Natl Acad Sci USA 74: 5463-5467). The sequence is given above. In a similar way, from a yeast line that, in spite of the his4/hipl double mutation, could be grown in a medium with histidine addition, the plasmid pFL61-aap2 was isolated whose insert was also cloned as a Notl fragment in pBluescriptSK. The resulting plasmid pAAP2 was sequenced and the sequence (Seq ID-No 2) is given above. The plasmid pAAP2 has a similar structure to pPPPl-20 (see Figure 1) , but instead of the insert Seq-ID No 1, carries the insert Seq-ID No 2 (see Figure 3) . Example 3 Uptake studies with 14 C-labelled protein into the yeast line PPP1-20 and AAP2 The yeast lines 22574d: :PPPl-20 and 22574d: :AAP2, that were obtained in a similar manner to Example 1, were grown in liquid medium until the culture reached the logarithmic phase. After centrifuging the culture, the cells are washed and taken up in 100 mm tris/HCI pH 4.5, 2mM MgCl 2and 0.6M sorbitol. Around 100 μl of the suspension was added to a solution of 0.5mM L-proline plus 1 μCi 14 C labelled L- proline in 100 μl of the same buffer. The uptake of the labelled amino acid was measured by the process described by Cirillo (1989, Meth Enzymol 174: 617-622) . The uptake of the labelled amino acid was compared, on the one hand in co-incubation with protein modifying substance diethyl pyrocarbonate, which is an inhibitor of the amino acid transport in membrane vesicles from Beta vulgaris, and on the other hand in co-incubation with other protein modifying substances. The calculated reduction is shown in Tables I and/or III. A competition experiment in which the specificity of the transporter could be read off with various amino acids and analogues is shown in Table II for PPP1-20 and in Figure 4 for AAP2. An analogous experiment in which a competition for histidine uptake in the line JT16::AAP2 was tested, is described in Example 5. The time period for PPP1-20 is shown in Figure 2. Example 4 Transformation of plants with a construct for over- expression of the coding region of amino acid transporters From the plasmid pPPPl-20, that contains as insert the cDNA for the amino acid transporter from Arabidopsis, an internal fragment of the insert was isolated after BamHI cleavage and cloned in the BamHI cleavage position from pAJ, that was first linearised with the enzyme BamHI. Then the cDNA was prepared as the EcoRI/HindHI fragment from pA7 and cloned in the vector pBIN-HYG. After transformation by Agrohacteria , this was inserted for infection of leaf segments of tobacco and potato. Ten independently obtained transformands, in which the presence of the intact non rearranged chimeric gene was demonstrated using Southern blot analysis, were tested for modifications of amino acid and nitrogen content. Besides this, amino acid synthesis, photosynthesis rate and transportation were tested. Example 5 Studies in the uptake of 1 C-labelled histidine in the yeast line AAP2 The yeast line JT16::AAP2, that was obtained in a similar manner to Example 1 was grown in liquid medium until the culture reached the logarithmic phase. After centrifuging the culture, the cells were washed and taken up in 10 mm tris/HCI pH 4.5, 2 mm MgCl 2 and 0.6M sorbitol. Around 100 ml of the suspension was added to a solution of 0.5 mm L- histidine plus 1 μCi 14 C-labelled L-histidine in 100 μl of the same buffer. The uptake of the labelled amino acid was measured according to the method described by von Cirillo (1989, Meth Enzymol 174: 617-622). The uptake of the labelled amino acid was compared in a competition experiment with that from different amino acids and analogues in tenfold excess. The relationships are shown in Figure 5. Table I Inhibition of the amino acid transport in 22574d: :PPP1- 20 - yeast strains by protein modifying substances % of transport without inhibitor 0.1 mM DEPC 65 (diethyl pyrocarbonate) 10 μM CCCP <3 (Carbonyl cyanide m-chlorophenylhydrazone) 10 μM 2,4 DNP <3 (Dinitrophenol) 1 mM sodium arsenate 35 10 μM antimycin A 29 500 μM PCMBS 78 (p-chloromercuribenzenesulfonic acid) Table II Competition by one, fourfold and tenfold excess of amino acids and analogues in 22574d: :PPPl-20 - veast strain 4 x lOx 27 30 27 83 79 58 88 74 50 21 14 95 49 48 45 77 47 67 57 27 18 21 8 16 29 23 71 48 16 44 90 Table III Inhibition of the amino acid transports in JT16::AAP2 - yeast strain by protein modifying substances % of transport without inhibitor 1 mM DEPC 3.1 ± 1.6 (Diethyl pyrocarbonate) 10 μM CCCP 15.6 ± 2.1 (Carbonyl cyanide m-chlorophenylhydrazone) 10 μM 2,4 DNP 7.6 ± 1.6 (Dinitrophenol) BUDAPESTTREATYONTHEINTERNATION AL RECOGNITIONOFTHEDEPOSITOFMICROORGANIS MS FORTHEPURPOSESOFPATENTPROCEDURE INTERNATIONALFORM Institut fur Genbiologische Forsc-aung Berlin GmbH Ihnestrasse 63 lOOO Berlin 33 VIABILITY STATEMENT issued puι*-ruant to Rule 10 E by the INTERNATIONAL DEPOSITARY AUTHORITY identified at the bottom of thin page I. DEPOSITOR II. IDENTIFICATION OF THE MICROORGANISM ame: Institut fύr Genbiologische Accession number given by the Fors chung Berlin GmbH INTERNATIONAL DEPOSITARY AUTHORITY: ddress: Ihnestrasse 63 lOOO Berlin 33 DSM 7129 Date of the deposit or of the transfer : 1992-06-23 HI. VIABILITY STATEMENT The viability of the microorganism identified under U above was tested on 1992 — 06—2 On that date, the said microorganism was ( X ) 3 viable ( ) no longer viable IV. CONDITIONS UNDER WHICH THE VIABILITY TEST HAS BEEN PERFORMED* IV. INTERNATIONAL DEPOSITARY AUTHORITY Name: DSM DEUTSCHE S M LUNG VON Signacure(s) of ρerson(s) having the power MIKROORGANISMEN UND ZELLKULTUREN GmbH to represent the International Depositary Authority Address: M&scheroder Weg 1 B or of authorised offϊcial(s): D-3300 Braunschweig Date: 1992-06-30 1 Indicate the date of original depo-.it or, where a new deposit or a transfer has been made, the most recent relevant date (date of the new deposit or date of the transfer). In the ca-sea referred to in Rule 10.2(a) (u) and (tit), refer to the most recent viability test. Mark with a cross the applicable box, Fill in if the information has been requested and if the results of the test were negat ή Form DSM-BP/9 (sole page) 0787 SUBSTITUTE SHEET BUDAPEST TREATY ON THE INTERNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FOR THE PURPOSES OF PATENT PROCEDURE INTERNATIONAL FORM Institut fur Genbiologische Forschung Berlin GmbH Ihnestrasse 63 lOOO Berlin 33 RECEIPT IN THE CASE OF AN ORIGINAL DEPOSIT issued pursuant to Rule 7.1 by the INTERNATIONAL DEPOSITARY AUTHORITY identified at the bottom of this page I. IDENTIFICATION OF THE MICROORGANISM Identification reference given by the DEPOSITOR Accession number given by the INTERNATIONAL DEPOSITARY AUTHORITY: pPPPl-20 DSM 7129 π. SCIENTIFIC DESCRIPTION AND/OR TAXONOMIC DESIGNATION The microorganism identified under I. above was accompanied by: ( X ) a scientific description ( } a proposed taxooomic designation (Mark with a cross where applicable) III. RECEIPT AND ACCEPTANCE This International Depositary Authority accepts this microorganism identified under I. above, which was received by it on 1992 -06 -23 (Date of original deposit) 1 IV. RECEIPT OF REQUEST FOR CONVERSION The microorganism identified under I above was received by this International Depositary Authority on (date of original deposit) and a request to convert the original deposit to a deposit under the Budapest Treaty was received by it on (date of receipt of request for conversion). V. INTERNATIONAL DEPOSITARY AUTHORITY Name: DSM-DEUTSCHE SAMMLUNG VON Signature(s) of person(s) having the power MIKROORGANISMEN UND ZELLKULTUREN GmbH to represent the International Depositary Authority or of authorized official(s): Adress: Mascheroder V/eg I 3 D-3300 Braunschweig Date: 1992-06-30 Where Rule 6.4(d) applies, such date is the date on which the status of international depositary auchority was acquired. Form DSM-BP/*. (sole page) 0291 SUBSTITUTE SHEET BUDAPEST TREATY ON THE INTERNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FOR THE PURPOSES OF PATENT PROCEDURE INTERNATIONAL FORM Institut fύr Gehbiologische Forschung Berlin GmbH Ihnestrasse 63 1000 Berlin 33 VIABILITY STATEMENT issued pursuant to Rule 10.2 by the INTERNATIONAL DEPOSITARY AUTHORITY identified at the bottom of this page I. DEPOSITOR π. IDENTIFICATION OF THE MICROORGANISM Name: Institut fύr Genbiologische Accession number given by the Forschung Berlin GmbH Addri Ihnestrasse 63 INTERNATIONAL DEPOSITARY AUTHORITY: 1000 Berlin 33 DSM 7130 Date of the deposit or of the traα j fer : 1992-06-23 HL VIABILITY STATEMENT The viability of the microorganism identified under II above was tested on 1992 —06 — 24 On that date, the said microorganism was ( X ) 3 viable ( ) no longer viable IV. CONDITIONS UNDER WHICH THE VIABILITY TEST HAS BEEN PERFORMED* IV. INTERNATIONAL DEPOSITARY AUTHORITY Name DSM DEUTSCHE SAMMLU G VON- Sιgnature(s) cl person(s) having :.**.e power MIKROORGANISMEN UND ZELL LTURSN GmbH to represent tne Intema:,onal Desoβitary Autnoπ t y Address: Mascheroder Weg 1 3 or of authorised officι--l(s): D-3300 Braunschweig Date: 1992-06-30 Indicate the date of original deposit or, wnere a new deposit or a tran s f er hu h .„ i . . . _ , ,_ , , H o r a t rans l er has been made, tne most recent relevant da t e (da t e of tae ae - d deeppoossiitt oorr ddaattee ooff tthhee ttrraannssffeerr)).. l In tne cases referred to in Rule 10.2(a) (,i) and (iii), refer to the most recent viabil i ty test Marx with a cross the applicable box. Fill i n i f the information has oeen requested and if the results of the test were negat i ve. Form DSM-BP/9 (sole page) 07S7 SUBSTITUTE SHEET BUDAPEST TREATY ON THE INTERNATIONAL RECOGNITION OF THE DEPOSIT OP MICROORGANISMS FOR THE PURPOSES OF PATENT PROCEDURE INTERNATIONAL FORM Institut fύr Genbiologische Forschung Berlin GmbH Ihnestrasse 63 1000 Berlin 33 RECEIPT IN THE CASE OF AN ORIGINAL DEPOSIT issued pursuant to Rule 7.1 by the INTERNATIONAL DEPOSITA-RY AUTHORITY identified at the bottom of thu page I. IDENTIFICATION OF THE MICROORGANISM Identification reference given by the DEPOSITOR Accession number given by the INTERNATIONAL DEPOSITARY AUTHORITY pBin PPP 1-20 DSM 7130 E. SCIENTIFIC DESCRIPTION AND/OR TAXONOMIC DESIGNATION The microorganism identified under I. above was accompanied by: ( X ) a scientific description ( ) a proposed taxonomic designation (Mark with a cross wnere applicable) IH. RECEIPT AND ACCEPTANCE This international Depositary Authoπty accepts this microorganism identified under I. aoove, which was received by it on 1992 -06-23 (Date of on-final deposit) 1 IV. RECEIPT OF REQUEST FOR CONVERSION The microorganism identified under I above was received by this international Depositary Autnonty on (date of original αepositl and a request to convert tne original deoosic to a deposit under the Buαaoest Treaty was i received by :t en (date cf recc:=: o. request for conversion) I V. INTERNATIONAL DEPOSITARY AUTHORITY I Name DSM-DEUTSCKE SAMMLUNG VON Signatured) ol person(s) navmg the power MIKROORGAN1SMEN UND ZELLKULTUREN GmbH to represent the international Depositary Autnonty or of authorised offιcιai(s) Mascneroder Weg 1 B D-2300 Braunschweig Date* 1992-06-30 * Where Ruie 5 *i(d) applies, suc αate is tne αate on w nicn the status of international Depositary au t hori t y was acquirec Form DSM-3P/4 (sole page) 0291

Claims: Claims 1. D N A sequences which contain the coding region of an amino acid transporter, characterised in that the information contained in the nucleotide sequence allows, by integration in a plant genome, the formation of RNA, and with this RNA, a new amino acid transport activity can be introduced in the plant cells or an endogenous amino acid transporter 10 activity can be expressed. A DNA sequence according to claim 1, characterised in that, it contains the following nucleotide sequence (Seq-ID No 1) : CTTAAAACAT TTATTTTATC TTCTTCTTGT TCTCTCTTTC TCTTTCTCTC ATCA C T 5 6 ATG AAG AGT TTC AAC ACA GAA GGA CAC AAC CAC TCC ACG GCG GA A 101 Met Lys Ser Phe Asn Thr Glu Gly His Asn His Ser Thr Ala Glu 1 5 10 15 TCC GGC GAT GCC TAC ACC GTG TCG GAC CCG ACA AAG AAC GTC GAT 145 Ser Gly Asp Ala Tyr Thr Val Ser Asp Pro Thr Lys Asn Val Asp 20 25 30 GAA GAT GGT CGA GAG AAC- CGT ACC GGG ACG TGC- CTT ACG GCG AGT 1 9 1 Glu Asp Gly Arg Glu Lys Arg Thr Gly Thr Trp Leu Thr Ala Ser 35 40 45 GCC- CAT ATT ATC ACG GCG GTG ATA GGC TCC GGA GTC- TTG TCT TTA 2 3 5 Ala His Ilε He Thr Ala Val He Gly Ser Gly Val Leu Ser Leu 50 55 60 GCA TGG GCT ATA GCT CAG CTT GGT TGG ATC GCA GGG ACA TCG ATC 281 Ala Trp Ala He Ala Gin Leu Gly Trp He Ala Gly Thr Ser He 65 70 75 TTA CTC ATT TTC TCG TTC ATT ACT TAC TTC ACC TCC ACC ATG CTT 325 Leu Leu He Phe Ser Phe He Thr Tyr Phe Thr Ser Thr Met Leu 80 85 90 GCC GAT TGC TAC CGT GCG CCG GAT CCC GTC ACC GGA AAA CGG AAT 371 Ala Asp Cys Tyr Arg Ala Pro Asp Pro Val Thr Gly Lys Arg Asn 95 100 105 TAC ACT TAC ATG GAC GTT GTT CGA TCT TAC CTC GGT GGT AGG AAA 415 Tyr Thr Tyr Met Asp Val Val Arg Ser Tyr Leu Gly Gly Arg Lys HO 115 120 GTG CAG CTC TGT GGA GTG GCA CAA TAT GGG AAT CTG ATT GGG GTC 461 Val Gin Leu Cys Gly Val Ala Gin Tyr Gly Asn Leu He Gly Val 125 130 135 ACT GTT GGT TAC ACC ATC ACT GCT TCT ATT AGT TTG GTA GCG GTA 505 Thr Val Gly Tyr Thr. He Thr Ala Ser He Ser Leu Val Ala Val ■ 14 ° 145 150 GGG AAA TCG AAC TGC TTC CAC GAT AAA GGG CAC ACT GCG GAT TGT 551 Gly Lys Ser Asn Cys Phe His Asp Lys Gly His Thr Ala Asp Cys 155 160 165 ACT ATA TCC- AAT TAT CCG TAT ATG GCG GTT TTT GGT ATC ATT CA.- "=-- Thr He Ser Asn Tyr Pro Tyr Met Ala Val Phe Gly He He Glr. 170 175 180 GTT ATT CTT AGC CAG ATC CCA AAT TTC CAC AAG CTC TCT TTT CT" 6- * .- Val He Leu Ser Gin He Pro Asn Phe His Lys Leu Ser Phe Leu 85 190 1SΞ TCC ATT ATG GCC GCA GTC ATG TCC TTT ACT TAT GCA ACT ATT GGA 686 Ser He Met Ala Ala Val Met Ser Phe Thr Tyr Ala Thr He Glv 200 205 210 ATC GGT CTA GCC ATC GCA ACC GTC GCA GGT GGG AAA GTG GGT AAG 731 He Gly Leu Ala He Ala Thr Val Ala Gly Gly Lys Val Gly Lys 215 220 225 ACG AGT ATG ACG GGC ACA GCG GTT GGA GTA GAT GTA ACC GCA GCT 776 Thr Ser Met Thr Gly Thr Ala Val Gly Val Asp Val Thr Ala Ala 230 235 240 CAA AAG ATA TGG AGA TCG TTT CAA GCG GTT GGG GAC ATA GCG TTC 82 _ Gin Lys He Trp Arg Ser Phe Gin Ala Val Gly Asp He Ala Phe 245 250 255 GCC TAT GCT TAT GCC ACG GTT CTC ATC GAG ATT CAG GAT ACA CTA 866 Ala Tyr Ala Tyr Ala Thr Val Leu He Glu He Gin Asp Thr Leu 260 265 270 AGA TCT AGC CCA GCT GAG AAC AAA GCC ATG AAA AGA GCA AGT CTT 911 Arg Ser Ser Pro Ala Glu Asn Lys Ala Met Lys Arg Ala Ser Leu 275 280 285 GTG GGA GTA TCA ACC ACC ACT TTT TTC TAC ATC TTA TGT GGA TG C 956 Val Gly Val Ser Thr Thr Thr Phe Phe Tyr He Leu Cys Gly Cys 290 295 300 ATC GGC TAT GCT GCA TTT GGA AAC AAT GCC CCT GGA GAT TTC CTC LOCI He Gly Tyr Ala Ala Phe Gly Asn Asn Ala Pro Gly Asp Phe Leu 305 310 315 ACA GAT TTC GGG TTT TTC GAG CCC TTT TGG CTC ATT GAC TTT GCA T 046 Thr Asp Phe Gly Phe Phe Glu Pro Phe Trp Leu He Asp Phe Ala 320 325 330 SUBSTITUTE SHEET AAC GCT TGC ATC GCT GTC CAC CTT ATT GGT GCC TAT CAG GTG TTC 1091 Asn Ala Cys He Ala Val His Leu He Gly Ala Tyr Gin Val Phe 335 340 345 GCG CAG CCG ATA TTC CAG TTT GTT GAG AAA AAA TGC AAC AGA AAC 1136 Ala Gin Pro He Phe Gin Phe Val Glu Lys Lys Cys Asn Arg Asn 350 355 360 TAT CCA GAC AAC AAG TTC ATC ACT TCT GAA TAT TCA GTA AAC GTA 1181 Tyr Pro Asp Asn Lys Phe He Thr Ser Glu Tyr Ser Val Asn Val 365 370 375 CCT TTC CTT GGA AAA TTC AAC ATT AGC CTC TTC AGA TTG GTG TGC- 1226 Pro Phe Leu Gly Lys Phe Asn He Ser Leu Phe Arg Leu Val Trp 380 385 390 AGG ACA GCT TAT GTG GTT ATA ACC ACT GTT GTA GCT ATG ATA TTC 1271 Arg Thr Ala Tyr Val Val He Thr Thr Val Val Ala Met He Phe 395 400 405 CCT TTC TTC AAC GCG ATC TTA GGT CTT ATC GGA GCA GCT TCC TTC 1316 Pro Phe Phe Asn Ala He Leu Gly Leu He Gly Ala Ala Ser Phe 410 415 420 TGG CCT TTA ACG GTT TAT TTC CCT GTG GAG ATG CAC ATT G C A CAA 1361 Trp Pro Leu Thr Val Tyr Phe Pro Val Glu Met His lie Ala " Gin 425 430 435 ACC AAG ATT AAC- AAG TAC TCT GCT AGA TGG ATT GCC- CTC- AAA ACC- 1405 Thr Lys He Lys Lys Tyr Ser Ala Arg Trp He Ala Leu Lys Thr 440 445 450 ATG TGC TAT GTT TGC TTG ATC GTC TCG CTC TTA GCT GCA GCC GGA 1451 Met Cys Tyr Val Cys Leu He Val Ser Leu Leu Ala Ala Ala Glv 455 460 465 SUBSTITUTE SHEET TCC ATC GCA GGA CTT ATA AGT AGT GTC AAA ACC TAC AAG CCC TTC 1496 Ser He Ala Gly Leu He Ser Ser Val Lys Thr Tyr Lys Pro Phe 470 475 480 CGG ACT ATG CAT GAG TGAGTTTGAG ATCCTCAAGA GAGTCAAAAA 1541 Arg Thr Met His Glu 485 TATATGTAGT AGTTTGGTCT TTCTGTTAAA CTATCTGGTG TCTAAATCCA 1591 ATGAGAATGC TTTATTGCTA AAACTTCATG AATCTCTCTG TATCTACATC 1641 TTTCAATCTA ATACATATGA GCTCTTCCAA AAAAAAAAAA AAAA 1685 3. A DNA sequence according to claim 1, characterised in that it contains the following nucleotide sequence (Seq-ID No 2) : CTATTTTAT AATTCCTCTT CTTTTTGTTC 29 ATAGCTTTGT AATTATAGTC TTATTTCTCT TTAAGGCTCA ATAAGAGGAG 79 AT G GGT GAA ACC GCT GCC GCC AAT AAC CAC CGT CAC CAC CAC CAT 124 Met Gly Glu Thr Ala Ala Ala Asn Asn His Arg His His His His 1 5 10 15 CAC GGC CAC CAG GTC TTT GAC GTG GCC AGC CAC GAT TTC GTC CCT i 5S His Gly His Gin Val Phe Asp Val. Ala Ser His Asp Phe Val Pro 0 25 30 CCA CAA CCG GCT TTT AAA TGC TTC GAT GAT GAT GGC CGC CTC AAA 21- Pro Gin Pro Ala Phe Lys Cys Phe Asp- Asp Asp Gly Arg Leu Lys 35 40 45 SUBSTITUTE S AGA ACT GGG ACT GTT TGG ACC GCG AGC GCT CAT ATA ATA ACT GCG 259 Arg Thr Gly Thr Val Trp Thr Ala Ser Ala His He He Thr Ala 50 55 60 GTT ATC GGA TCC GGC GTT TTG TCA TTG GCG TGG GCG ATT GCA CAG 304 Val He Gly Ser Gly Val Leu Ser Leu Ala Trp Ala He Ala Gin 65 70 75 CTC GGA TGG ATC GCT GGC CCT GCT GTG ATG CTA TTG TTC TCT CTT 349 Leu Gly Trp He Ala Gly Pro Ala Val Met Leu Leu Phe Ser Leu 80 85 90 GTT ACT CTT TAC TCC TCC ACA CTT CTT AGC GAC TGC TAC AGA ACC 394 Val Thr Leu Tyr Ser Ser Thr Leu Leu Ser Asp Cys Tyr Arg Thr 5 100 105 GGC GAT GCA GTG TCT GGC AAG AGA AAC TAC ACT TAC ATG GAT GCC 439 Gly Asp Ala Val Ser Gly Lys Arg Asn Tyr Thr Tyr Met Asp Ala HO 115 120 GTT CGA TCA ATT CTC GGT GGG TTC AAG TTC AAG ATT TGT GGG TT G 484 Val Arg Ser He Leu Gly Gly Phe Lys Phe Lys He Cys Gly Leu 125 130 135 ATT CAA TAC TTG AAT CTC TTT GGT ATC GCA ATT GGA TAC ACG ATA 529 He Gin Tyr Leu Asn Leu Phe Gly He Ala He Gly Tyr Thr He 140 145 150 GCA GCT TCC ATA AGC ATG ATG GCG ATC AAG AGA TCC AAC TGC TTC 57-. Ala Ala Ser He Ser Met Met Ala He Lys Arg Ser Asn Cys Phe 155 160 165 CAC AAG AGT GGA GGA AAA GAC CCA TGT CAC ATG TCC AGT AAT CCT 61= His Lys Ser Gly Gly Lys Asp Pro Cys His Met Ser Ser Asn Pro 70 175 130 SUBSTITUTE SHEET TAC ATG ATC GTA TTT GGT GTG GCA GAG ATC TTG CTC TCT CAG GTT 664 Tyr Met He Val Phe Gly Val Ala Glu He Leu Leu Ser Gin Val 185 190 200 CCT GAT TTC GAT CAG ATT TGG TGG ATC TCC ATT GTT GCA * GCT GTT 709 Pro Asp Phe Asp Gin He Trp Trp He Ser He Val Ala Ala Val 205 210 220 ATG TCC TTC ACT TAC TCT GCC ATT GGT CTA GCT CTT GGA ATC GTT 754 Met Ser Phe Thr Tyr Ser Ala He Gly Leu Ala Leu Gly He Val 225 230 235 CAA GTT GCA GCG AAT GGA GTT TTC AAA GGA AGT CTC ACT GGA ATA 799 Gin Val Ala Ala Asn Gly Val Phe Lys Gly Ser Leu Thr Gly He 240 245 250 AGC ATC GGA ACA GTG ACT CAA ACA CAG AAG ATA TGG AGA ACC TTC 844 Ser He Gly Thr Val Thr Gin Thr Gin Lys He Trp Arg Thr Phe 255 260 265 CAA GCA CTT GGA GAC ATT GCC TTT GCG TAC TCA TAC TCT GTT GTC 889 Gin Ala Leu Gly Asp He Ala Phe Ala Tyr Ser Tyr Ser Val Val 270 275 280 CTA ATC GAG ATT CAG GAT ACT GTA AGA TCC CCA CCG GCG GAA TCC- 934 Leu He Glu He Gin Asp Thr Val Arg Ser Pro Pro Ala Glu Ser 285 290 295 AAA ACC- ATG AAG AAA GCA ACA AAA ATC AGT ATT GCC GTC ACA ACT 97S Lys Thr Met Lys Lys Ala Thr Lys He Ser He Ala Val Thr Thr 300 305 310 ATC TTC TAC ATG CTA TGT GGC TCA ATG GGT TAT GCC GCT TTT GGA 1024 He Phe Tyr Met Leu Cys Gly Ser Met Gly Tyr Ala Ala Phe Gly 315 320 325 SUBSTITUTE SHEET GAT GCA GCA CCG GGA AAC CTC CTC ACC GGT TTT GGA TTC TAC AAC 1069 Asp Ala Ala Pro Gly Asn Leu Leu Thr Gly Phe Gly Phe Tyr Asn 330 335 340 CCG TTT TGG CTC CTT GAC ATA GCT AAC GCC GCC ATT GTT GTC CAC 1114 Pro Phe Trp Leu Leu Asp He Ala Asn Ala Ala He Val Val His 245 350 355 CTC GTT GGA GCT TAC CAA GTC TTT GCT CAG CCC ATC TTT GCC TTT 1159 Leu Val Gly Ala Tyr Gin Val Phe Ala Gin Pro He Phe Ala Phe 360 365 370 ATT GAA AAA TCA GTC GCA GAG AGA TAT CCA GAC AAT GAC TTC CTC 1204 He Glu Lys Ser Val Ala Glu Arg Tyr Pro Asp Asn Asp Phe Leu 375 380 385 AGC AAG GAA TTT GAA ATC AGA ATC CCC GGA TTT AAG TCT CCT TAC 1249 Ser Lys Glu Phe Glu He Arg He Pro Gly Phe Lys Ser Pro Tyr 390 395 400 AAA GTA AAC GTT TTC AGG ATG GTT TAC AGG AGT GGC TTT GTC GTT 1294 Lys Val Asn Val Phe Arg Met Val Tyr Arg Ser Gly Phe Val Val 405 410 415 ACA ACC ACC GTG ATA TCG ATG CTG ATG CCG TTT TTT AAC GAC GTG 1339 Thr Thr Thr Val He Ser Met Leu Met Pro Phe Phe Asn Asp Val 420 425 430 GTC GGG ATC TTA GGG GCG TTA GGG TTT TGG CCC TTG ACG GTT TAT 13S Val Gly He Leu Gly Ala Leu Gly Phe Trp Pro Leu Thr Val Tyr 435 440 445 TTT CCG GTG GAG ATG TAT ATT AAG CAG AGG AAG GTT GAG AAA TGG 142S Phe Pro Val Glu Met Tyr He Lys Gin Arg Lys Val Glu Lys Trp 450 455 460 SUBSTITUTE SHEET AGC ACG AGA TGG GTG TGT TTA CAG ATG CTT AGT GTT GCT TGT CTT 1474 Ser Thr Arg Trp Val Cys Leu Gin Met Leu Ser Val Ala Cys Leu 465 470 475 GTG ATC TCG GTG GTC GCC GGG GTT GGA TCA ATC GCC GGA GTG ATG 1519 Val He Ser Val Val Ala Gly Val Gly Ser He Ala Gly Val Met 480 485 490 CTT GAT CTT AAG GTC TAT AAG CCA TTC AAG TCT ACA TAT 1558 Leu Asp Leu Lys Val Tyr Lys Pro Phe Lys Ser Thr Tyr 495 500 TGATGATTAT GGACCATGAA CAACAGAGAG AGTTGGTGTG TAAAGTTTAC 1608 CATTTCAAAG AAAACTCCAA AAATGTGTAT ATTGTATGTT GTTCTCATTT 1653 CGTATGGTCT CATCTTTGTA ATAAAATTTA AAACTTATGT TATAAATTAT 1708 AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AA 1 740 4. A plasmid, characterised in that it contains a DNA sequence according to any one of claims 1 to 3. 5. Plasmid pPPPl-20 (DSM 7129) . 5 6. Plasmid pAAP2, prepared according to Example 2. 7. Plasmid pBin PPP1-20 (DSM 7130) . 10 8. Use of the plasmid according to any one of claims 4 to 7 or derivatives or parts thereof, for the transformation of prokaryotic and eukaryotic cells. 9. Plants containing a DNA sequence according to any one 15 of claims 1 to 3. SUBSTITUTE SHEET 1 0 . Bacteria containing a DNA sequence according to any one of claims 1 to 3. 11. Use of the DNA sequence of the amino acid transporter according to any one of claims 1 to 3 for the preparation of plasmids with changed specificity of the transporter. 12. Use of the DNA sequence of the amino acid transporter according to any one of claims 1 to 3 for isolation of similar sequences from the genome of the plant. 13. Use of the DNA sequence of the amino acid transporter according to any one of claims l to 3 for the expression of translatable mRNA, that makes possible the synthesis of an amino acid transporter in prokaryotic and eukaryotic cells. 14. Use of the DNA sequence of the amino acid transporter according to any one of claims 1 to 3 for the expression of a non-translatable mRNA, that hinders the synthesis of an amino acid transporter in prokaryotic and eukaryotic cells. 15. Use of the DNA sequence of the amino acid transporter according to any one of claims 1 to 3 in combination with steering elements for an expression in prokaryotic and eukaryotic cells. 1 6. Yeast strains containing DNA sequences according to any one of claims 1 to 3. 1 7. Use of yeast strains containing DNA sequences according to claim 16 for identification of a plant amino acid transporter. 18. Use of the DNA sequence of the amino acid transporter according to any one of claims 1 to 3 for preparation of plants with changed amino acid and nitrogen metabolism. 19 . Use of the DNA sequence of the amino acid transporter according to any one of claims 1 to 3 for preparation of crop plants with increased yield. 2 0. Use of the DNA sequence of the amino acid transporter according to any one of claims 1 to 3 for the transport of compounds in prokaryotic and eukaryotic cells.

Inventors: FROMMER WOLF-BERND, FROMMER, WOLF-BERND
Applicants: FROMMER WOLF BERND, INST GENBIOLOGISCHE FORSCHUNG, INSTITUT FUER GENBIOLOGISCHE FORSCHUNG BERLIN GMBH
Application year: 1993
Main classification: C12N15
IPC codes: A01H5, C07K14, C12N1, C12N5, C12N15