摘要
为了研究蚯蚓金属硫蛋白的生物学功能,采用RT-PCR方法从镉诱导后的赤子爱胜蚓(Eisenia foetida)中克隆到蚯蚓金属硫蛋白基因cDNA。将其克隆入原核表达载体pET-DsbA中,然后转化至大肠杆菌表达受体菌BL21(DE3)中进行表达。产物经SDS-PAGE进行分析,可见约32.4 kD的融合蛋白。Zn2+抗性测定表明含pET-DsbA-efMT的重组菌较含空载体pET-DsbA的对照菌重金属抗性有所提高。
To study the function of Eisenia foetida metallothionein, the cDNA fragment encoding the Eisenia foetida metallothionein was amplified by RT-PCR from the Eisenia foetida induced by cadmium and cloned into prokaryotic expression vector pET-DsbA. After transformation of the recombinant plasmid into expression host Escherichia coli BL21（DE3）, the expression product was analyzed by SDS-PAGE electrophoresis, and an approximately 32.4 kDa fusion protein was detected. Zinc ion resistance assay showed that the recombinant strain exhibited increased heavy metal resistance compared with non-recombinant control E. coll.
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2010年第1期84-87,92,共5页
Journal of Hebei Agricultural University
基金
河北省教育厅项目(2007412)
关键词
MT基因
克隆
原核表达
抗性
metallothionein gene
clone
prokaryotic expression
resistance
