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Position: Home > Articles > Cloning and Prokaryotic Expression 1D Gene of YC2011 Strain of Porcine Sapelovirus Progress in Veterinary Medicine 2014,35 (1) 72-76

猪萨佩罗病毒YC2011株1D基因的克隆及原核表达

作  者:
陈俊伟;张祥斌;张云静;周庆丰;宋延华;李薇;陈峰;薛春宜;毕英佐
单  位:
华南农业大学动物科学学院;广东温氏食品集团股份有限公司;中山大学生命科学学院
关键词:
猪萨佩罗病毒;1D基因;原核表达
摘  要:
为研发猪萨佩罗病毒(PSV)检测试剂,根据PSV YC2011毒株核苷酸序列,设计针对1D基因的特异引物,以YC2011毒株为模板,利用RT-PCR方法,成功扩增出1D基因,将该基因与原核表达载体pET-32a(+)连接,转化表达菌株BL21。经PCR和测序鉴定,阳性菌株IPTG诱导表达,融合蛋白1D进行SDS-PAGE和Western blot检测分析。结果表明,成功构建了原核表达菌株,其所表达的融合蛋白分子质量约为51ku,且可被猪萨佩罗病毒阳性血清所识别。
译  名:
Cloning and Prokaryotic Expression 1D Gene of YC2011 Strain of Porcine Sapelovirus
作  者:
CHEN Jun-wei;ZHANG Xiang-bin;ZHANG Yun-jing;ZHOU Qing-feng;SONG Yan-hua;LI Wei;CHEN Feng;XUE Chun-yi;BI Ying-zuo;CAO Yong-chang;Guangdong Wen's Foodstuffs Group Co.Ltd.;College of Animal Science,South China Agricultural University;State Key Laboratory of Biocontrol,School of Life Sciences,Sun Yat-sen University;
关键词:
Porcine sapelovirus;;1D Gene;;eucaryotic expression
摘  要:
According to the porcine Sapelovirus YC2011strain nucleotide sequence reported in the GenBank, we designed a pair of primers to amplify 1Dgene of YC2011strain.The 1Dgene was cloned into prokaryotic expression vector pET-32a(+)and transformed into E.coli strain BL21.In order to obtain 1Dprotein, the positive strains were induced by IPTG,and then the expressed products were analyzed by SDS-PAGE and Western blot.The results of SDS-PAGE and Western blot demonstrated that the 1Dgene of YC2011 strain was successfully expressed.The expressed 1Dprotein has 51ku,and can be recognized by the positive serum of Porcine sapelovirus.

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