摘要
为了建立肌钙蛋白-Ⅰ(troponinⅠ,TnⅠ)的原核表达,试验以人肝脏cDNA文库为模板,通过PCR方法扩增出肌钙蛋白-Ⅰ基因,将其克隆到PCR-TOPO质粒中,构建表达质粒pET-22b(+)-TnⅠ,转化大肠杆菌BL21(DE3)-RPX并进行诱导表达。结果表明:表达的目的蛋白占菌体总蛋白的20%以上,并主要以可溶形式存在。说明试验建立了troponinⅠ的非亲和色谱纯化工艺,重组蛋白具有良好的抗血管生成活性。
To establish the prokaryotic expression of troponin I , the human troponin I gene was obtained by PCR amplification using a human liver cDNA library as a template. This gene was cloned into PCR -TOPO vector to construct expression plasmid pET- 22b（ ＋ ） -Tn I , and then the plasmid was transformed into E. coli BL21 （ DE3 ） - RPX and induced the expression. The results showed that the expressed target protein accounted for more than 20% of the total bacterial proteins, and existed mainly in soluble form. The result demonstrates that the non - affinity chromatography purification of troponin I is constructed, and the recombinant protein has good anti - angiogenic activity.
出处
《黑龙江畜牧兽医》
CAS
北大核心
2013年第7期13-16,共4页
Heilongjiang Animal Science And veterinary Medicine
关键词
肌钙蛋白-Ⅰ
表达
纯化
抗血管生成
troponin I
expression
purification
anti - angiogcnesis
