外源Toll样受体2转基因绵羊的构建与鉴定

Toll样受体2（Toll-likereceptor2，TLR2）是识别革兰氏阳性细菌如引起羊乳腺炎、炭疽病、破伤风等病原体的一种重要受体，而TLR2过表达转基因羊为研究TLR2在防卫细菌侵染中的作用提供了一个理想的模型。因此，研究首先通过显微注射TLR-2过表达载体3S—Loxp—TLR2的方法对绵羊进行外源基因TLR2整合，然后采用PCR和Southernblot方法对其进行鉴定、拷贝数验证和表达量分析。结果表明，注射148个受精卵，共产下15只小羊，其中8只小羊检测为阳性，转基因率为53．3％；Southern杂交结果发现转TLR2小羊的基因组中外源TLR2基因均为单拷贝；且转基因羊TLR2mRNA表达高于对照。说明成功构建了转TLR2基因羊。

Construction and Identification of Transgenic Toll-like Receptor 2 Sheep

TLR2 （Toll-like Receptor 2） was the important receptor to recognize the invaded gram-positive microbes which caused serious mastiffs, anthrax,tetanus and so on. Transgenic sheep constitutively over-expressing TLR2 in all tissues would be the ideal model to uncover the role of TLR2 on bacteria clearance. TLR-2 over-expression vector （3S-Loxp-TLR2） was used to generate transgenic sheep by egg microinjection. One hundred and forty eight zygotes was microinjected TLR-2 over-expression vector and fifteen lambs were born. The results showed that totally 8 transgenic sheep were identified by PCR and southern blot analysis,the integration efficiency of foreign gene by nficroinjection reached 53.3%. We also found exogenous TLR2 was one copy in all transgenic sheep. The expression level of TLR2 gene of different transgenic lambs were analyzed by using real time PCR, the results showed that transgenic sheep built higher levels of mRNA expression and protein secretion of TLR2.