摘要
从含有目的基因的cDNA克隆中,利用PCR方法钓取目的基因.将目的基因与酶切线性化的载体进行定向连接,其产物转化细菌感受态细胞.采用菌落PCR方法和核酸序列测定进行阳性克隆鉴定.对构建好的pCMV-tag2A/NAP1融合蛋白表达载体进行超纯去内毒素抽提.
NAP1 gene CDS was amplified from pMD-18T/NAP1 vector by using PCR technique. The purified PCR products and pCMV-tag2A vector were ligated and transformed into host strain E.coil DH5a. Clones containing the vectors were selected on LB-plus ampicillin（100~tg/ml） plates, and plasmid DNA was extracted and digested with enzymes. Recombinant expression plasmids pCMV-tag2A/NAP1 were constructed from pMD-18T/NAP1 vector and pCMV- tag2A expression vector. Plasmids of containing the right insertion were sequenced to confirm its identify and retransformed into E.coil DH5a. Finally, pCMV-tag2A/NAP 1 expression vector were performanced endotoxin extraction.
出处
《湖南文理学院学报（自然科学版）》
CAS
2011年第4期53-56,共4页
Journal of Hunan University of Arts and Science(Science and Technology)
基金
湖南省自然科学基金项目(08JJ3037)
湖南省高校创新平台开放基金项目(09K097)
湖南省高校动物学重点实验室资助
湖南省大学生研究性创新性学习与创新性实验项目资助
