摘要
[目的]克隆大鼠神经营养因子BDNF基因,构建植物表达载体,在豌豆植物中表达BDNF蛋白。[方法]采用RT-PCR法克隆大鼠脑源性神经营养因子BDNF基因,构建豌豆植物表达载体p CAPE2-BDNF,利用豌豆发芽种子真空侵染法在豌豆植物中瞬时表达BDNF蛋白,以His标签抗体进行Western Blot检测目的蛋白。[结果]获得含有鼠源性神经营养因子BDNF基因的植物表达载体p CAPE2-BDNF,His标签抗体检测到目的条带。[结论]BDNF蛋白在豌豆植物中成功表达,有助于进一步对其功能活性进行分析。
[Objective]Brain derived neurotrophic factor（ BDNF） of rat was amplified and constructed into the pea plants expression vector p CAPE2. BDNF proteins were expressed in pea plants. [Methods]After cloned by RT- PCR method,the BDNF gene was constructed into pea plants expression vector p CAPE2,and the p CAPE2- BDNF vectors were introduced into the pea plants via the vacuum infiltration of germinated seeds method,the anti- His polyclonal antibody was used to detect the recombinant protein of BDNF. [Results]Pea plant expression vector p CAPE2- BDNF was constructed; the recombinant proteins were confirmed by blotting with anti- His polyclonal antibody. [Conclusion]BDNF protein expressed in pea plants successfully which helped to analyze the function of BDNF in vitro.
出处
《生物技术》
CAS
CSCD
北大核心
2016年第2期136-139,共4页
Biotechnology
基金
国家自然基金项目(No.31350003)
吉林省科技厅项目(No.20130102063JC)
长春师范大学自然科学基金(No.2011D010)
