Application of a wap gene in Drosophila transgenic
A transgenic, fruit fly technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problems of incalculable social significance and economic value of antimicrobial peptide drugs, low yield and activity, insoluble recombinant protein, etc. Antibacterial function and viability, significant antibacterial effect, high expression effect
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[0043] The present invention will be further described below.
[0044] (1) Construction of WAP transgenic fruit flies: According to the sequence at both ends of the ORF of the antibacterial peptide gene WAP containing a single whey acidic protein domain in Penaeus chinensis, a pair of upstream and downstream primers Wap-f and Wap-r were designed, and the upstream and downstream The 5' ends of the primers were introduced separately kpn I(GGTACC) with xba I (AGATCA) restriction site, the primer sequence is as follows:
[0045] Wap-f: 5'---GG GGTACC ATGGTGAACATCAAGGAAGTTC---3' (SEQ ID NO: 1)
[0046] Wap-r:5'---GCTCT AGATCA T TTTCCGTAGGGAGATCCCA ---3' (SEQ ID NO: 2)
[0047] Using the cDNA of hemocytes of Penaeus chinensis as a template, the WAP fragment was caught by PCR, and the kpn I and xba ⅠTwo restriction sites were directionally connected with the pUAST plasmid to construct the eukaryotic expression vector wap / pUAST, and the sequence of the inserted WAP fragme...
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