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CVN(Cyanovirin-N)是一种高效的抗HIV蛋白。Cyanovirin-N(CVNH)家族是CVN的同源蛋白,具有与CVN的肽链折叠类型并且具有高度保守的抗HIV的结构域。把水蕨Ceratopteris thalictroides野生型和突变型CtCVNH蛋白的编码序列分别克隆到真核表达载体pP ICZαA,结果所有野生型wC tC VNH和突变型mCtCVNH基因分别产生两条带,Western blot确认大小分别为22 000和24 000。采用超滤离心和Ni-NTA亲和色谱法纯化蛋白质,糖基化检测证实wC tC VNH蛋白不存在N-糖基化位点,可能的原因是载体pPICZαA中α因子信号肽序列的不完全切割或存在一些非糖基化的其它翻译后修饰,为今后进一步研究CtCVNH的药代动力学和药效学奠定了坚实的基础。
CVN (Cyanovirin-N) is a highly effective anti-HIV protein. The Cyanovirin-N (CVNH) family is a cognate protein of CVN that has a type of peptide chain folding with CVN and has a highly conserved anti-HIV domain. The coding sequences of both wild-type and mutant CtCVNH proteins of Ceratopteris thalictroides were cloned into eukaryotic expression vector pP ICZαA respectively. As a result, all wild-type wC tC VNH and mutant mCtCVNH genes produced two bands, respectively, and the sizes were 22 000 and 24 000. The protein was purified by ultrafiltration and Ni-NTA affinity chromatography. The glycosylation assay confirmed the absence of N-glycosylation sites in the wC tC VNH protein, possibly due to incomplete cleavage of the alpha factor signal peptide sequence in the vector pPICZαA Or some other non-glycosylated post-translational modifications, which laid a solid foundation for further research on the pharmacokinetics and pharmacodynamics of CtCVNH in the future.