植物病毒沉默抑制子P25的原核表达及纯化  

Expression of A RNA Silencing Suppressor in Eschcrichia coli and Its Purification

作　　者：秦波[1] 麻鹏达[1] 伏秀青[1] 杨涛[1] 刘晶莹[1] 董春光[1] 王兴智[1] 

机构地区：[1]东北师范大学遗传与细胞研究所,长春130024

出　　处：《吉林农业大学学报》2005年第4期378-380,共3页Journal of Jilin Agricultural University

基　　金：国家自然科学基金资助项目(30370767)

摘　　要：通过体外DNA重组技术将P25基因连接在pET28a(+)载体上,构建成重组质粒。经PCR扩增、酶切鉴定及DNA序列分析,融合的P25基因序列正确。将重组质粒转化到大肠杆菌BL21(DE3)中进行异丙基-βD硫代半乳糖苷(IPTG)诱导表达,经过亲和层析得到纯品目的蛋白P25。The P25 gene was inserted into the pET - 28a（ ＋ ） vector to create a recombinant plasmid pET - 28a（ ＋ ） - P25. The resulted recombinant DNA was verified by PCR, restriction enzyme digestion and DNA sequencing. The P25 protein was then expressed in Eschcrichia coli and purified by Ni-NTA affinity chromatography after being induced by IPTG under denaturing condition.

关 键 词：基因沉默抑制子 P25蛋白 原核表达系统 

分 类 号：Q786[生物学—分子生物学]