| 摘要: |
| 为克隆滩羊肝脏α-生育酚转移蛋白(α-TTP)CDS区基因并构建其原核表达载体,通过Trizol法提取滩羊肝脏组织总RNA,将其反转录为cDNA后利用人工合成和PCR扩增相结合的方法得到α-TTP CDS区基因,并将其克隆至原核表达载体pET28a,命名为pET28a-TTPA。结果表明:扩增了滩羊肝脏α-TTP CDS区基因,同已公布的绵羊α-TTP序列同源性达99.76%,并且将其成功克隆至原核表达载体pET28a。结果为后续α-TTP基因原核表达及其抗体的制备奠定了基础。 |
| 关键词: 滩羊 α-生育酚转移蛋白 基因克隆 原核表达载体 |
| DOI：10.11841/j.issn.1007-4333.2012.04.018 |
| 投稿时间：2012-02-13 |
| 基金项目:国家自然科学基金资助项目(31172230) |
|
| Cloning of Tan Sheep liver alpha TTP gene and its original nuclear expression vector construction |
|
JIA Hui-na, LUO Hai-ling, LIU Kun, ZUO Zhao-yun, ZHANG Yu-wei, WANG Zhen-zhen
|
| (State of Key Laboratory of Animal Nutrition/College of Animal Science and Technology, China Agricultural University, Beijing 100193, China) |
| Abstract: |
| The aim of this study was to clone the CDS area of α-TTP of Tan Sheep and constructed its original nuclear expression vector.RNA of theTan Sheep liver was extracted through Trizol method,and it was reverse transcripted to cDNA.The CDS area of α-TTP was got through synthetic method and PCR amplify method,and it was eventually cloned to original nuclear expression vector pET28a,and named pET28a-TTPA.The results showed that,the CDS area of α-TTP was not only amplified and cloned to the original nuclear expression vector pET28a successfully,but alsoshowed a 99.76% sequence homology with the published sheep α-TTP gene.These results established the foundation for α-TTP prokaryotic expression and the preparation of its antibody. |
| Key words: Tan Sheep α-tocopherol transfer protein gene cloning original nuclear expression vector |
