%0 Journal Article
%T Cloning and Prokaryotic Expression of Human Recombinant Calreticulin<br>重组人钙网蛋白的克隆与原核表达
%A CAO Chun-yu
%A HAN Yu
%A WANG Yan-lin
%A <br>曹春雨
%A 韩钰
%A 王艳林
%J 中国生物工程杂志
%D 2008
%I 
%X Objective: Clone, express and purify human recombinant calreticulin (CRT). Methods: Human CRT cDNA was amplified by RT-PCR from total RNA of human lung cancer cell line A549 cells. Then, PCR product was subcloned into prokaryotic expression vector pET-15b. After sequencing, this recombinant plasmid was transformed into E. coli. Rossetta. Recombinant CRT was expressed in host cells by IPTG induction. Resulted protein was purified by Ni-NTA resin under denature condition and dialyzed to recover its native structure. SDS-PAGE and Western blot method were used to identify the expression and purification of reconbinant CRT. Results: Human CRT cDNA was cloned from total RNA of A549 cells. CRT prokaryotic expression vector pET-15b-crt was constructed. Reconbinant CRT was induced to express in E. coli and purified by Ni-NTA affinity chromatograph. Conclusion: A method for prokaryotic expression and purification of human recombinant CRT was successfully established. This method laid a foundation for the succedent CRT research.
%K calreticulin
%K prokaryotic expression
%K protein purification<br>钙网蛋白
%K 原核表达
%K 蛋白纯化
%K 重组人
%K 钙网蛋白
%K 克隆与原核表达
%K Recombinant
%K Human
%K Prokaryotic
%K Expression
%K 功能研究
%K 实验方法
%K 高度
%K 诱导性
%K 重组质粒
%K 结果
%K 状态
%K Western
%K blotting
%K 透析复性
%K 纯化
%K 亲和层析
%K 树脂
%K 条件
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=951380788B355DEA7D75520FA3B199C9&aid=DEECC90EDCA0F01F690E2D256C14A752&yid=67289AFF6305E306&vid=D3E34374A0D77D7F&iid=E158A972A605785F&sid=09ABD5535D9B6D45&eid=B9704B40A4225A24&journal_id=1671-8135&journal_name=中国生物工程杂志&referenced_num=1&reference_num=8