期刊论文

中文

现代生物医学进展

1673-6273

2008

8

3

449-452

10.3969/j.issn.1673-6273.2008.03.015

湖南省自然科学基金项目（20050203）；

本校为第一机构

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 dye...