禽流感抗原基因NA，HA的克隆及其表达载体的构建  

Cloning of NA and HA Gene Fragments from Avian Influenza Virus and Constructing of NA and HA Expression Vector

作　　者：李进[1] 阮颖[1] 龚剑[2] 范亚丽[1] 姚远頲 杜培粉[1] 魏然[3] 刘春林[3] 

机构地区：[1]湖南农业大学生物科学技术学院,长沙410128 [2]中国科学院病毒生物所,武汉430070 [3]湖南农业大学作物基因工程湖南省重点实验室,长沙410128

出　　处：《现代生物医学进展》2008年第3期449-452,共4页Progress in Modern Biomedicine

基　　金：湖南省自然科学基金项目(20050203)

摘　　要：HA和NA是禽流感病毒重要的保护性抗原基因,为了得到禽流感植物疫苗,本试验采用高保真PCR扩增方法得到目的基因,分别克隆到pMD18-T载体。经测序证实核酸序列正确后,克隆到含有GUS基因的高效植物双元表达载体pBl121上,获得含有HA/NA基因的植物双元表达载体pBI121-HA和pBI121-NA,采用冻融法将含HA/NA基因的植物双元表达载体转入根癌农杆菌LBA4404,菌液浸染生菜子叶,共培48小时后进行GUS基因表达检测,x-glue染色显蓝色,说明带有HA/NA的植物双元表达载体构建成功,为下一步的生菜转HA/NA基因研究奠定基础。HA and NA are two of important antigens of Avian influenza virus. In order to obtain Avian influenza virus plant vaccine, Target genes HA and NA were amplified by high-fidelity PCR and cloned into the transition pMD18-T vector. After sequencing, HA and NA were ligated into the plant binary vector pBI 121 respectively. New plant binary vectors, named pBI 121-HA and pBI 121-NA, were constructed, and than transferred into Agrobacterium LBA4404. Excise dcotyledons were inoculated by immersion and after co-cultivation for 48 hours the expression of GUS in cotyledon was detected. Cotyledons were dyed by x-gluc. The plant binary expression vector encoding NA/HA had been constructed successfully, and facilitated fulther investigation of NA/HA protein expressions in Lactuca.

关 键 词：禽流感抗原基因NA HA 植物高效表达载体 禽流感 

分 类 号：Q819[生物学—生物工程]