Spreadsheet: 6d0767ac-0f08-48ab-be19-fdec16842af1
Sheet: Sheet1

Table S5: Primer Information. F: forward primer; R: reverse primer. | Unnamed: 1 | Unnamed: 2
Primer | Sequence | Note
[NAME] | GGTCTAGAACACAGTGGAAAAAATCGACAT | For transgenic constructs of kep1; pro: promoter plus 5' UTR; CDS: coding sequences
[NAME] | GGGAATTCAACAATCTTTACAGAAAATTAC
[NAME] | GGGAATTCATGATAAAAATGGAAACCCCAA
[NAME] | GGGAATTCTTTATATTTTTTAAGTATTGTATACG
nsr/CG3927-pro-F | GGTCTAGAACATAATGGGCAACCGACATT | For transgenic constructs of nsr; pro: promoter plus 5' UTR; CDS: coding sequences. The primers for amplifing promoter region are the same between nsr and CG3927. The required PCR products were confirmed by sequencing.
nsr/CG3927-pro-R | GGGAATTCTTTTATCACAATAATCTTTACCG
nsr-CDS-F | GGGAATTCATGGAAACCCAAAGCGAGTTT
nsr-CDS-R | GGGAATTCATTATAGCGTTGTGCGACGAA
nsr/CG3927-pro-F | GGTCTAGAACATAATGGGCAACCGACATT | For transgenic constructs of CG3927; pro: promoter plus 5' UTR; CDS: coding sequences. The primers for amplification promoter region are the same between nsr and CG3927. The required PCR products were confirmed by sequencing.
nsr/CG3927-pro-R | GGGAATTCTTTTATCACAATAATCTTTACCG
CG3927-CDS-F | GGGAATTCATGGAAACCCCAAGCGAGTT
CG3927-CDS-R | GGGAATTCTAGACGGACGACTGGGGAG
CG4021-pro-F | GGTCTAGAACATAATGGAAAACCGATACTT | For transgenic constructs of CG4021; pro: promoter plus 5' UTR; CDS: coding sequences
CG4021-pro-R | GGGAATTCTTTTTCCATAACAATCTTTAGC
CG4021-CDS-F | [NAME]
CG4021-CDS-R | GGGAATTCCTGTGCTCCAAGCATAGATTT
[NAME]F | GGTCTAGAACATGGGAAAATATTGAAATTTA | For transgenic constructs of D. yakuba [NAME]; pro: promoter plus 5' UTR; CDS: coding sequences
[NAME] | GGGAATTCTTTTAACATAACACTTCTAAC
[NAME] | GGGAATTCATGGAGACCCCAAGCGAGTTTA
[NAME] | GGGAATTCTTTATATTTTTTGAGTATGGAATA
GFP-F | GGGAATTCATGGTGAGCAAGGGCGAGGAG | For amplifing GFP of transgenic constructs
GFP-R | GGGGTACCTTACTTGTACAGCTCGTCCATGC
TAP-F | GGGAATTCACTAGTTCCATGGAAAAGAGAAGATG | For amplifing TAP of transgenic constructs
TAP-R | GTGGTACCACTAGTTCAGGTTGACTTCCCCGCGGAGTT
nsr-1 | TCGAGCTCGCGGCCGCATTAAGTTAGTGCATGGAAATCA | nsr gene targeting construct: 1. The construct is designed to delete the entire coding region of nsr from the start codon to a point downstream of the stop codon.  2. The PCR product from primers nsr-3 and nsr-4 was ligated into the pBS vector using the SalI site and the NotI site. 3. An I-SceI site was then inserted at the NotI site by ligating in the double stranded oligonucleotide produced by annealing of the oligonucleotides I-SceI-Not1 and I-SceI-Not2. 4. The PCR product of primers nsr-1 and nsr-2 was then added to the plasmid at the SacI site. Then clone with right direction was selected. 5. DNA segments between primers nsr-5 and nsr-6 was then amplified and ligated into the plasmid using the SalI and XhoI sites. 6. The assembled construct was released from that vector using the NotI sites and transferred into the same site of pTV2.
nsr-2 | TCGAGCTC TGTTTTGTGCACCTAATTAA
nsr-3 | [NAME]
nsr-4 | CCGTCGACATTTTTATCACAATAATCTTTACC
nsr-5 | GCGTCGACACAAAGGACAATGCTTACGTTG
[NAME] | CAA CTCGAGGCGGCCGCTGCCTCTTGTCCACTTTATCCTG
I-SceI-Not1 | GGCCTAGGGATAACAGGGTAAT
I-SceI-Not2 | GGCCATTACCCTGTTATCCCTA
CG4021-1 | TCGAGCTCGCGGCCGCAGGAGAATAATGAATATTCAAATG | CG4021 gene targeting construct: 1. The construct is designed to delete the entire coding region of CG4021, from the start codon to a point downstream of the stop codon  2. The PCR product from primers CG4021-3 and CG4021-4 was ligated into the pBS vector using the XbaI site and the NotI site. 3. An I-SceI site was then inserted at the NotI site by ligating in the double stranded oligonucleotide produced by annealing of the oligonucleotides I-SceI-Not1 and I-SceI-Not2. 4. The PCR product of primers CG4021-1 and CG4021-2 was then added to the plasmid at the SacI site. Clone with right direction of insertion was selected. 5. DNA segments between primers CG4021-5 and CG4021-6 was then amplified and ligated into the plasmid using the XbaI and XhoI sites. 6. The assembled construct was released from that vector using the NotI sites and transferred into the same site of pTV2.
CG4021-2 | TCGAGCTCTTCAATAGCTCGGCATGTGCCC
CG4021-3 | GGTAAATGCGGCCGCGAACAACCATCGACCGGGATA
CG4021-4 | CCTCTAGA ACAATCTTTAGCGAAAAATCACAAT
CG4021-5 | GCTCTAGAATGATACGTATACCCAAAATAATAA
CG4021-6 | CAAC/TCGAGCGGCCGCAATTATAACGAGCTGTCTAAGTG
I-SceI-Not1 | GGCCTAGGGATAACAGGGTAAT
I-SceI-Not2 | GGCCATTACCCTGTTATCCCTA
nsr-Rescue-F | GCGGCCGCTGTCTGATCTCCGCCAGAAC | For amplifing rescue fragment of nsr
nsr-Rescue-R | GCGGCCGCATCTTAGCTCGTTCTGAATTCGA
dj-promoter-F | GGTCTAGACGTCTTGGGTTATGGTCCTA | For amplifing promoter region of dj
dj-promoter-R | GGTCTAGAGAGTTCACTGCATACCTTCC
[NAME] | CGGGATCCATCAACCTGATTAAGAGAGCTG | The resulted PCR product of Kep1 antigen was cloned into PGEX.
[NAME] | GCCTCGAGAAGTATTGTATACGGATTGGG
[NAME] | GTTCACTTCGACCGGATAGAAT | Real-time PCR primers: for detecting mature transcript
[NAME]-R | GGCGGATCAACATATCGAG
[NAME] | GCCTATATGGGTCATTACGAT
[NAME]-R | CCTTGCCTCTTCTCAACATAAG
kl-5-F | CTATACCTTCGACGGATGGAG
kl-5-R | ATCCTATCACTTGGCAATCCTT
kl-2-pre-F | TATATGTGCATATATCTCCAGCAT | Real-time PCR primers: for detecting pre-mRNA. The primers are designed to amplify the exon-intron junction.
kl-2-pre-R | CCCAACTATTAAAATGTCCTACA
kl-3-pre-F | TTCCGTTTTTATTTACAATTTT
kl-3-pre-R | GAGATCCATATTATATCCTCGTAT
kl-5-pre-F | TTTGGTCCGATTTTTAGATCACT
kl-5-pre-R | TCCATGAAAAGACCGTTGATATAG
rp49-F | GCTAAGCTGTCGCACAAATGGC | Real-time internal control
rp49-R | GCGCTTGTTCGATCCGTAACC
[NAME] | AGTTTACTGAGAAACAGCCACCC | For RT-PCR
[NAME] | GGTGTGCGACGAAATCAACC
nsr-F | CCAAGTTCAACTTCGCTCGG
nsr-R | TGCGACGAAATCACTTGGC
CG3927-F | CATTCCCGGCAAGGAGCC
CG3927-R | GTCCACATAGCAGGATTGGGC
CG4021-F | AGATTACTGAGGAACAGCCAACT
CG4021-R | TTCATAACGTAGGTGCCTATACC
rp49-F | GCTAAGCTGTCGCACAAATGGC
rp49-R | GCGCTTGTTCGATCCGTAACC