人C6orf210基因原核表达载体的构建及蛋白的初步表达

为了构建人C6orf210基因原核表达载体，并在E．coliBL21中表达并纯化。采用RT—PCR扩增人C6orf210基因片段，并将其克隆到原核表达载体Pet21a中，构建重组质粒pET21a—C6ort210。经限制性内切酶EcoRI与XhoI双酶切鉴定及序列测定后，转化E．coliBL21，经IPTG诱导表达融合蛋白。结果显示获得全长为1188bp的人的CAiorf210基因片段。以构建的重组质粒pET21a—C6orf210转化E．coliBL21后，经IPTG诱导，表达出相对分子质量（Mr）约66000的融合蛋白。经SINS—PAGE、Western blot分析显示，诱导表达的蛋白为C6orf210。

Construction of the Prokaryotic Expression Vector and Expression of Human C6orf210 Gene

This paper tries to construct human C.6orf210 gene and express the gene in E. coli BL21 and purities. RT-PCR is used to expands increase the human C6orf210 gene fragment, and clones it to the nucleus expresses in carrier Pet21a, to construct reorganization material particle pET21a - C6orf210. IPTG induction expression fusion protein. Results show that the span is approximately 1188 bp person＇s C6off210 gene fragment. After pet21a- C6off210 transforms E. coli BL21, IPTG induction, expression showing off to molecular mass （Mr） approximately 66000 fusion proteins.