大鲵虹彩病毒贵州分离株MCP基因的克隆与原核表达
发布时间：2019-07-13 21:42
【摘要】：根据Gen Bank中大鲵虹彩病毒主衣壳蛋白MCP(major capsid protein,MCP)基因序列(序列号:KF512820),设计一对特异性引物,以大鲵虹彩病毒贵州分离株基因组DNA为模板,PCR扩增大鲵虹彩病毒MCP基因并测序,与Gen Bank中大鲵虹彩病毒MCP基因进行比对,然后将其亚克隆到原核表达载体p ET-32a(+)中,转化大肠杆菌BL21(DE3)感受态细胞,经IPTG诱导后进行Western blot分析。结果显示:PCR扩增出长度为1 392 bp的片段,与Gen Bank中大鲵虹彩病毒MCP基因核苷酸序列相似性为99.7%~99.9%,SDSPAGE电泳显示该重组蛋白的相对分子质量约为67×103。免疫原性检测结果表明,该重组蛋白可与兔抗大鲵虹彩病毒阳性血清特异性反应,具有免疫原性。
[Abstract]:According to the sequence (serial number: KF512820) of giant salamander rainbow virus main shell protein MCP (major capsid protein,MCP gene in Gen Bank, a pair of specific primers were designed. Using the genomic DNA of giant salamander rainbow virus Guizhou isolate as template, the MCP gene of giant salamander rainbow virus was amplified and sequenced by PCR, and compared with the MCP gene of giant salamander iridovirus in Gen Bank, and then subcloned into prokaryotic expression vector p ET-32a (). E. coli BL21 (DE3) receptive cells were transformed and induced by IPTG for Western blot analysis. The results showed that the fragment of 1392 bp was amplified by PCR, which was 99.7% 鈮，
本文编号：2514309
[Abstract]:According to the sequence (serial number: KF512820) of giant salamander rainbow virus main shell protein MCP (major capsid protein,MCP gene in Gen Bank, a pair of specific primers were designed. Using the genomic DNA of giant salamander rainbow virus Guizhou isolate as template, the MCP gene of giant salamander rainbow virus was amplified and sequenced by PCR, and compared with the MCP gene of giant salamander iridovirus in Gen Bank, and then subcloned into prokaryotic expression vector p ET-32a (). E. coli BL21 (DE3) receptive cells were transformed and induced by IPTG for Western blot analysis. The results showed that the fragment of 1392 bp was amplified by PCR, which was 99.7% 鈮，
本文编号：2514309
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