摘要
将水稻偏好密码子优化并人工合成抗虫基因cry1Ca通过限制性内切酶酶切的方法构建原核表达载体pET-28bca,成功表达了带6个组氨酸标签的His-cry1Ca融合蛋白,其表达量约占菌体总蛋白的24%,表达的融合蛋白主要以包涵体的形式存在。对包涵体蛋白进行可溶性处理,亲和纯化出带6个组氨酸标签的融合蛋白,将其作为抗原免疫家兔获得了滴度(ELISA)为l∶50 000的抗血清,并具有较强的特异性。
Rice codon optimized cry1Ca gene was cloned into expression vector pET-28b（＋） by means of restricted enzymatic digestion.His-cry1Ca fusion protein was highly expressed by SDS-PAGE analysis accumulating more than 24% of the total bacterial proteins.The main expression production was deposited as inclusion body.His-cry1Ca was then purified with Ni-NTA after treatment of solvents.The target protein was used as the antigen to immunize rabbits.ELISA assay showed that the titer of the prepared polyclonal antibody was 1：50 000,and had high specificities.
出处
《福建农业学报》
CAS
2011年第1期24-28,共5页
Fujian Journal of Agricultural Sciences
基金
福建省属公益类科研院所基本科研专项(2009R10035-3
2010R1021-3)
福建省科技计划重点项目(2007Y0057)
