期刊论文

中文

中国病原生物学杂志

1673-5234

2015

10

9

796-800

10.13350/j.cjpb.150907

特殊病原体防控湖南省重点实验室资助项目（湘科计字[2014]5号,湘教通〔2012〕312号）；

本校为第一机构

目的构建解脲支原体Ferritin蛋白的原核表达载体,纯化重组Ferritin蛋白并测定其抗氧化功能,为解析解脲支原体的抗氧化机制奠定基础。方法利用荧光定量PCR检测氧化胁迫下Ferritin基因的相对表达量。根据Ferritin基因序列,设计引物,PCR扩增Ferritin全长片段。将解脲支原体Ferritin基因中编码色氨酸的密码子TGA突变为TGG,将突变后的Ferritin基因连接到原核表达载pET28a得到重组质粒pET28a-Ferritin,转化大肠埃希菌BL21(DE3)得到重组菌BL/pET28a-Ferritin,IPTG诱导Ferritin蛋白表达并用镍柱亲和层析进行纯化。体外试验测定Ferritin蛋白的Fe~(2+)结合特性,亚铁氧化酶活性和抗氧化能力。结果Ferritin基...

Objectives The aims of this study were to clone a ferritin gene from Ureaplasma urealyticum, to over-ex- presses and purify the recombinant ferritin protein in E. coli, and to evaluate the anti-oxidative activity of ferritin. The protective role of ferritin should provide a foundation for understanding the mechanisms by which U. urealyticurn tolerates oxidative stress. Methods The relative level of ferritin expression under oxidative stresses was analyzed using realtime PCR. The ferritin gene from U. urealyticum was cloned using PCR. The geneti...