转rhG-CSF基因鱼腥藻的重组基因及蛋白检测  

Identification of Recombinant Gene and Protein of Transgenic Anabaena sp. 7120-G-CSF

作　　者：赵兴贵[1] 李清艳[2] 李爽[1] 施定基[1] 

机构地区：[1]天津科技大学海洋科学与工程学院,天津300457 [2]南开大学生命科学学院,天津300071

出　　处：《海洋通报》2007年第5期108-111,共4页Marine Science Bulletin

基　　金：天津市科技攻关重点资助项目(No.043182711);天津科技大学自然科学基金资助项目(No.20050208)

摘　　要：提取质粒进行PCR和酶切鉴定人粒细胞集落刺激因子(rhG-CSF)基因成功转入鱼腥藻7120中,并用酶联免疫方法测得转基因鱼腥藻7120-G-CSF中G-CSF蛋白在第21d表达量达到最大。对转基因蓝藻的生长曲线、叶绿素a含量测定结果表明,rhG-CSF基因转化蓝藻后,对蓝藻前中期的生长影响不大,但21d后生长速度下降,同时外源蛋白表达也达到最大值,这有利于转基因藻的大规模培养。After being transferred into Anabaena sp. PCC 7120, the vector pRL-G-CSF in transgenic Anabaena 7120-G-CSF was confirmed by PCR and restriction analysis, and the expression efficiency of protein G-CSF reached the maximum accumulation which was measured using ELISA. The result of assaying the growth curve, content of chlorophyll indicated that the growth of transgenic Anabaena wasn＇t affected so much, but the cell numbers of transgenic Anabaena reduced in culturing 21 days in liquid medium, at the same time, the expression of protein G-CSF reached the maximum accumulation, which are helpful to culture transgenic Anabaena in scale.

关 键 词：人粒细胞集落刺激因子 PCR 酶切 转基因鱼腥藻7120-G—CSF G-CSF蛋白表达量 

分 类 号：Q784[生物学—分子生物学]