新疆南疆微小牛蜱Bm86/Bm91双基因原核表达载体的构建

为构建微小牛蜱Bm86和Bm91双基因原核表达载体,以新疆生产建设兵团塔里木畜牧科技重点实验室获得的Bm86和Bm91克隆基因阳性菌或质粒DNA为模板,参照GenBank登录的微小牛蜱Bm86和Bm91基因序列和所用载体序列,设计可以扩增Bm86和Bm91基因完整阅读框架的引物,然后对新疆南疆微小牛蜱Bm86和Bm91基因进行PCR扩增、纯化、酶切后连接,构建双基因重组质粒pET-32a-Bm并鉴定。结果显示,成功构建Bm86和Bm91双基因重组质粒pET-32a-Bm,经测序鉴定和初始获得的Bm86和Bm91基因序列一致,未发生变异。

Construction of Prokaryotic Expression Vector of Bm86 and Bm91 Dual Genes of Boophilus microplus in South Xinjiang

In order to construct the prokaryotic expression vector of Bm86 and Bm91 dual genes of Boophilus microplus in South Xinjiang, the primers that can link, transform and amplify complete open reading frame of Bm86 and Bm91 dual genes were designed by reference to the sequences of genes and vectors of Bm86 and Bm91 of Boophilus microplus in GenBank. With the bacteria or plasmid DNA containing Bm86 and Bm91 has been gained in our laboratory as a template, the Bm86 and Bm91 of Boophilus microplus in South Xinjiang were amplified by PCR and connected. The recombinant plasmid pET-32a-Bm containing Bm86 and Bm91dual genes were constructed and identified. The full length and sequence of nucleotide acid of the inserted genes in extracted recombinant plasmid were completely identical to the Bm86 and Bm91.