摘要
应用RT-PCR方法,克隆394 bp的MTP片段,连接到pMD 18-T载体上构建pMD-MTP394重组质粒;应用限制性内切酶ApaⅠ消化重组质粒pMD-MTP394,切下一194 bp的小片段,回收大的线性片段,T4 DNA连接酶连接,成功构建了一个重组的竞争模板质粒pMD-MTP200,为建立竞争性RT-PCR法检测乳牛肝MTP基因mRNA奠定了基础。
The objective of the present study was to construct a recombinant DNA fragment which can be used as a competitive template for detecting changes of MTP mRNA in liver of dairy cows, A 194 bp fragment was cut from a 394 bp MTP recombinant plasmid by restriction enzyme Apa Ⅰ . Then, the larger linear fragment was ligated by T4 DNA ligase. The developed recombinant competitive template plasmid could be applied for quantification of MTP mRNA in liver of dairy cows.
出处
《中国兽医科技》
CAS
CSCD
北大核心
2005年第9期731-734,共4页
Chinese Journal of Veterinary Science and Technology
基金
国家自然科学基金项目(30230260)
