大鯢虹彩病毒貴州分離株MCP基因的克隆與原核表達
發(fā)布時間：2019-07-13 21:42
【摘要】：根據(jù)Gen Bank中大鯢虹彩病毒主衣殼蛋白MCP(major capsid protein,MCP)基因序列(序列號:KF512820),設(shè)計一對特異性引物,以大鯢虹彩病毒貴州分離株基因組DNA為模板,PCR擴增大鯢虹彩病毒MCP基因并測序,與Gen Bank中大鯢虹彩病毒MCP基因進行比對,然后將其亞克隆到原核表達載體p ET-32a(+)中,轉(zhuǎn)化大腸桿菌BL21(DE3)感受態(tài)細胞,經(jīng)IPTG誘導(dǎo)后進行Western blot分析。結(jié)果顯示:PCR擴增出長度為1 392 bp的片段,與Gen Bank中大鯢虹彩病毒MCP基因核苷酸序列相似性為99.7%~99.9%,SDSPAGE電泳顯示該重組蛋白的相對分子質(zhì)量約為67×103。免疫原性檢測結(jié)果表明,該重組蛋白可與兔抗大鯢虹彩病毒陽性血清特異性反應(yīng),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，
本文編號：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，
本文編號：2514309
本文鏈接：http://www.sikaile.net/kejilunwen/jiyingongcheng/2514309.html
最近更新
教材專著
