人转化生长因子TGF-β_3基因合成及其在大肠杆菌中的表达

Expression of Human Transforming Growth Factor β_3 in E.coli BL21

目的 :构建能高效表达人转化生长因子 β3 (TGF β3 )的基因工程菌。方法 :用PCR方法扩增人转化生长因子 β3 的cDNA ,将该cDNA片段插入表达载体pET3c的表达框架中 ,构建了表达菌株BL2 1/pET TGF β3 。IPTG诱导表达。通过阳离子交换、变性分子筛层析纯化重组蛋白。用Western blot杂交检测重组蛋白的免疫原性。结果 :所构建的表达菌株 ,其TGF β3 的表达量占菌体总蛋白 2 5 %以上。经纯化获得了银染一条带的重组蛋白。免疫学检测表明重组蛋白具有较强的抗原特异性。结论 :人转化生长因子 β3 在大肠杆菌中实现了高效表达 ,获得了具有免疫原性的重组TGF β3 。

AIM:High-level expression system of human Transforming Growth Factor β 3 was constructed. METHOD:TGF-β 3 cDNA produced by PCR was cloned into the expression vector pET3c. The expression was induced by IPTG. The expressed TGF-β 3 was purified by ionic exchange and molecular sieve chromatography from the inclusion body of bacteria lysate. RESULT:The expression level of the recombinant TGF-β 3 in E. coli BL21 was about 25%. The purified TGF-β 3 has strong antigen specificity. CONCLUSION:Recombinant TGF-β 3 was produced with high yield in E.coli BL21.