人TLR4及MD2基因反义真核表达载体的构建

Construction of antisense expression vectors of human TLR4 and MD2

目的 构建人TLR4及MD2基因反义真核表达载体 ,为研究其在肺炎症反应中的作用奠定基础。方法 通过PCR技术扩增出TLR4及MD2基因片段 ,然后分别反向插入真核表达载体pEFBOS的多克隆位点 ,最后对产生的重组子进行酶切和测序鉴定。结果 扩增出的TLR4目的片段为 2 6kb ,MD2为 0 5kb ,并成功地连接到pEFBOS载体上 ,DNA测序结果也显示插入片段为目的片段。

Objective To construct antisense expression vectors of human TLR4 and MD2. Methods Partial sequences of TLR4 and MD2 gene were cloned by PCR method. The cloned fragments were reversely inserted into the multiclone sites of plasmid pEFBOS, respectively. The constructed recombinants were identified with enzyme digestion and DNA sequencing. Results The cloned fragments of TLR4 and MD2 were 2.6 kb and 0.5 kb, respectively, and successfully bound to pEFBOS. The inserted fragments were also proved to be the objective ones by DNA sequencing. Conclusion Antisense expression vectors of TLR4 and MD2 are successfully constructed.