中南医学科学杂志2016，Vol.44Issue(5)：494-498,5.DOI:10.15972/j.cnki.43-1509/r.2016.05.004
肌酐水解酶基因工程菌的构建及功能研究
The Construction of the Creatininase Gene Engineering Bacteria and Function Study
摘要
Abstract
Objective To construct recombinant plasmids of Creatininase, cloning into E. coli, and study its expression and enzyme activity. Methods According to the gene sequence of creatininase in Genbank(D45424.1),it was biosynthesized to get the gene fragment of creatininase C,after the PCR amplified,and then was connected with GV296 to construct the recombinant plasmid GV296￣C,and transformed to E.coli BL21(DE3) to construct gene engineering strain GV296￣C/BL21 (DE3).After SDS￣PAGE,the concentration of creatinine in the culture fluid was determined. Results The recombinant plasmid GV296￣C was constructed successfully,enzyme fragment was about 0.783Kb as electrophoresed shown.The sequencing result was consistent with gene order(D45424.1).The reconstructed plasmid GV296￣C were trans￣formed into E.col BL21(DE3) competent cells.The protein of molecular weight was approximately 29KD by SDS￣PAGE. Conclusions The engineering strain GV296￣C/BL21 ( DE3) is constructed successfully,and the expression of the creati￣ninase is induced to be efficient,which has the enzymes activity.关键词
肌酐水解酶/基因工程菌/肌酐/慢性肾功能衰竭Key words
creatininase/gene engineering bacteria/creatinine/chronic renal failure分类
医药卫生引用本文复制引用
杨波,蒋芬,朱艳,杨成,向铁勇,段绍斌,蒋云生..肌酐水解酶基因工程菌的构建及功能研究[J].中南医学科学杂志,2016,44(5):494-498,5.基金项目
湖南省自然科学基金项目( No:13JJ3082) （ No:13JJ3082）
