减蛋综合征病毒六邻体蛋白基因的克隆与真核表达载体的构建

应用降落PCR技术扩增出减蛋综合征病毒六邻体蛋白基因并将其克隆至pMDT-18载体上,酶切、PCR及测序结果表明插入的片段为目的基因.切下该目的基因定向克隆至pcDNA 3质粒构建六邻体蛋白基因真核表达载体pcDNA 3-hexon,经各种酶切、PCR鉴定及进一步测序鉴定,证明六邻体蛋白基因片段所插入的位置、大小、核苷酸序列和阅读框架正确无误,从而为下一步的转染、表达及进一步阐明EDSV六邻体基因结构与功能的关系和减蛋综合征基因工程苗的研究奠定了良好的基础.

Cloning of Hexon-encoding Gene of Egg Drop Syndrome Virus and Construction of Eukaryotic Expression Plasmids of Hexon Gene

Hexon protein gene of egg drop syndrome virus(EDSV) was amplified by touchdown PCR(TD-PCR) and cloned into pMD 18-T vector,and then sequencing results showed that the insert fragment was the target hexon gene of EDSV.The hexon gene was inserted into pcDNA 3 vector,resulting in the construction of pcDNA 3-hexon eukaryotic expression plasmids.The pcDNA 3-hexon recombinant plasmids were identified by PCR and digested with enzymes and sequenced to confirm its rightness.The results will lay a sound foundation for further expression and illustration of the relationship between the structure and function of hexon gene and the genic engineering vaccine of EDSV-gDNA.