【摘要】 本研究根据 Gen Bank发表的 GPV B株全基因核苷酸序列 ,设计了 1对用以扩增 GPV主要非结构蛋白 NS1基因的引物。通过 PCR技术 ,扩增出 NS1完整基因片段 1.9kb。将 PCR产物克隆到 p MD 18- T载体后进行序列测定及分析 ,结果表明 :GPV H1株 NS1基因全长 1884 bp,编码 6 2 7个氨基酸 ,与国外已发表的 B株核苷酸序列同源性为 98.15 %。同时将该目的基因正向插入到 GST融合蛋白原核表达载体 PGEX- 6 P- 1的 Bam H 多克隆位点 ,构建了 GPV NS1的原核表达载体 ,成功的表达了约 97KD的 NS1融合蛋白。更多还原

【Abstract】 According to the published sequence of GPV B strain in GenBank, a pair of primers was designed by Oligo4.0. NS1 gene of GPV was amplified by PCR. The completegene was cloned into the vector PMD18-T and sequenced. The result of sequence showed that NS1 gene comprised of 1884 its could able to encode 627 amnio acids. The cloned NS1 gene shared 98.15% homology with the B strain. Meanwhile,the NS1 gene was inserted in the BamHⅠ site of plasmid PGEX-6P-1 to construct prokaryotic expression vector of GPV NS1 gene,which expressed 97 kD fusion protein of NS1.更多还原