摘要
通过RT-PCR技术从一份猪粪中扩增并克隆了戊型肝炎病毒主要结构基因ORF2部分片段,大小为681 bp,将其克隆于pMD18-T载体。序列分析表明,与GenBank公布的序列一致。然后,将该基因亚克隆到植物表达载体p35S-2300-two-T-DNA-4×Ta1上,并将载体p2300上的CaMV35S启动子替换为E12启动子,构建成植物表达载体p2300-ORF2-E12。通过冻融法将重组质粒导入根癌农杆菌LBA4404中,获得了携带ORF2基因的根癌农杆菌菌株,为转基因植物工作奠定了基础。
The swine hepatitis E virus （SHEV）ORF2 fragment was amplified by RTPCR and cloned into pMD18-T vector. Sequence analysis showed that it consisted of 681 nucleotides and was identical to that of GenBank reported. Then the ORF＇2 gene was linked to the p35S-2300-two-T-DNA-4X Ta 1 vector on which the CaMV35S promoter was replaced by the inducible promoter El2. The plant expression vector p2300-ORF2-E12 was constructed and transited into Agrobacterium LBA440 By freeze-thaw method. All the experiments have laid the foundation for the future research on transgenic plant vaccine.
出处
《黑龙江农业科学》
2009年第4期90-94,共5页
Heilongjiang Agricultural Sciences
基金
黑龙江省博士后启动基金资助项目(2005037187)
