抗艾滋病病毒蛋白CVNH在大肠杆菌中表达条件的优化

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。

Optimization for Culture Condition of CVNH in Escherichia coli,A Novel Anti-HIV (Human Immunodeficiency Virus) Protein

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, A_600nm 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 A_600nm 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.