【摘要】 CVNH(Cyanovirin-N homology)蛋白家族是具有抗HIV活性的蓝藻抗病毒蛋白-N(Cyanovirin-N,CVN)的同源物。在前期研究中,已首次获得水蕨Ceratopteris thalictroides CVNH基因的全长序列,构建了pET32a-CtCVNH的原核表达载体。以此为基础,对CVNH在大肠杆菌Rosetta 2(DE3)中的表达条件进行优化。分别探讨了最佳培养基类型及成分、培养基初始pH、诱导时机、诱导剂浓度及诱导时间。优化后的诱导条件是:含24 g.L-1酵母提取物和72 g.L-1胰蛋白胨的TB培养基、培养基初始pH为8.0、菌液A600nm为0.6时加入1.6 g.L-1乳糖诱导6 h。CVNH蛋白的表达量较优化前提高了1.9倍。这为进一步开展CVNH的药物研制和开发奠定基础。更多还原

【Abstract】 CVNH(cyanovirin-N homology,CVNH) is the homology protein of CVN,which is a novel anti-HIV glycoprotein.In the previous study,we have firstly reported the full-length sequences of CVNH gene cloned from Ceratopteris thalictroides,and constructed prokaryotic expression plasmid pET32a-CtCVNH.The plasmid DNA was transformed into Rosetta 2(DE3) E.coli cells for expression of the recombinant protein.In the study,we optimized culture conditions of CVNH protein expression,including the types and ingredients of the culture medium,initial pH,A600nm value,induction agents and concentrations,induction time,and expression time,respectively.The optimized culture conditions contained TB medium with 24 g · L-1 yeast extract and 72 g · L-1 tryptone,initial pH 8.0,and lactose induced for 6 hours at A600nm 0.6,which would increase CVNH productivity to 1.9 times than before.This result laid a foundation to explore the drug development of CNVN.更多还原