禽流感病毒血凝素基因在烟草叶片中瞬时表达

Transient expression of hemagglutinin of avian influenza virus in the leaves of Nicotiana benthamiana

血凝素(Hemagglutinin,HA)蛋白是禽流感病毒(Avian Influenza Virus,AIV)最主要的保护性抗原,它不仅可以诱导产生特异性的抗体,还可以刺激机体产生细胞毒性T淋巴细胞(Cytotoxic lymphocyte,CTL)反应。探索禽流感ha在植物中的表达,提高禽流感ha的表达量,对于研制禽流感亚单位疫苗具有重要意义。主要进行了ha的密码子偏好优化,载体构建和烟草中瞬时表达,最终在烟草叶片RNA水平上检测到ha的转录,并通过Western Blot方法检测到目的蛋白的表达。此外,添加Kozak序列和删除载体上的除草剂抗性基因(Bialaphos resistance gene,bar)能提高ha的翻译水平。因此,烟草作为植物生物反应器表达HA具有重要的应用前景。

Hemagglutinin （HA） is the main protective antigen of avian influenza virus. It can not only induce the production of specif-ic antibodies, but also stimulate the response of cytotoxic lymphocyte （CTL） in the body. It is important for the manufacture of avain influenza subunit vaccines to explore the expression of ha in plants and improve the expression quantity of ha. This study focused on the optimization of the codons, the construction of vectors and the transient expression in Nicotianabenthamiana. Lastly, the expres-sion of the ha on the transcriptional level was detected by RT-PCR and the translational level by Western blotting assay. Besides this, the addition of Kozak sequence and the deletion of the bialaphos resistance gene （bar） could improve the translation level of the ha. So, Nicotianabenthamianahas an important application prospect to express the HA as a plant bioreactor.