番木瓜环斑病毒外壳蛋白基因的构建

CONSTRUCTION OF PAPAYA RINGSPOT VIRUS CUPSID PROTEIN GENE

以番木瓜环斑病毒黄斑分离物(PRV—YS)RNA为模板,在人工合成的两端引物引导下,反转录合成了外壳蛋白(CP)基因cDNA第一链并通过PCR扩增获得双链cDNA。用重组有cDNA的pUC19转化E.coli DH52,采用双脱氧链终止法进行了cDNA序列分析,结果表明CP基因为861nt,与文献报道的相比,同源率90%,且少连续的6nt。

In leading of the two terminal primers, the first strand of Cp gene cDNA was reverse transcripted from PRV-YS RNA, and amplified to get doublestrand cDNA through PCR. E.coli DH5α was transformed by pUC19 recombined with the cDNA.The whole CP gene was sequenced with dideoxy-mediated chain-termination method.The result shows that PRV CP gene has 861 nt, with 86nt different from the one reported, and 6nt in success less than the latteer