摘要
为获得ALAS的重组表达蛋白,以人肝脏组织cDNA为模板,通过PCR扩增得到ALAS全长片段并测序,成功构建了pET30a(+)-ALAS融合表达载体并转化大肠杆菌Rosetta(DE).IPTG诱导融合蛋白表达.SDS变性凝胶电泳结果显示,融合蛋白的分子量约70kDa,并且在上清液中有少量表达.经镍柱一步纯化得到His6-ALAS融合蛋白.
To express and purifyδ—aminolevulinic acid （ALAS）in E.coli,liver RNA was reverse tran—scribed into cDNA.The ALAS gene was amplified by PCR and confirmed by sequencing.The pET30a（＋）—ALAS recombinant vector was successfully constructed.The recombinant vector was transformed into Rosetta （DE3 ）and induced by IPTG.SDS—PAGE analysis showed that the fusion protein was about 70 kDa,and a small portion of the protein was expressed in the supernatant.The expressed His6—tagged pro—tein was purified by Ni2＋ affinity chromatography column and characterized by SDS—PAGE and confirmed by western blot analysis.
出处
《内蒙古师范大学学报（自然科学汉文版）》
CAS
北大核心
2014年第6期766-770,共5页
Journal of Inner Mongolia Normal University(Natural Science Edition)
基金
内蒙古自然科学基金资助项目(2014MS08110)
