草鱼生长激素在毕赤酵母中的高效分泌表达  

Expression of Grass Carp Growth Hormone in the Yeast Pichia pastoris

作　　者：王伟[1] 孙永华[1] 汪亚平[1] 朱作言[1] 

机构地区：[1]中国科学院水生生物研究所淡水生态与生物技术国家重点实验室,武汉430072

出　　处：《Acta Genetica Sinica》2003年第4期301-306,共6页

基　　金：国家自然科学基金项目 (3 0 12 3 0 0 4;90 2 0 80 2 4);国家 973项目 (G2 0 0 0 0 1610 9)资助~~

摘　　要：为大量获得具有生物学活性的草鱼生长激素 ,对草鱼生长激素cDNA在毕赤酵母中的表达进行了研究。将草鱼生长激素cDNA克隆入毕赤酵母表达载体pGAPZ α B ,构建表达载体pGAPZ α B GH。在三磷酸甘油醛脱氢酶 (GAP)启动子的调控作用下 ,一个类似于天然生长激素大小、分子量约 2 2kD的蛋白获得表达 ,其表达量约 5 0mg L。Western杂交表明 :表达的蛋白与兔抗草鱼生长激素的多克隆抗体特异结合 ,证实该表达蛋白为草鱼生长激素 ;受体夹心式ELISA检测表明 :表达的草鱼生长激素具生物学活性 。Growth hormone plays an essential role in the stimulation of somatic growth and development.To produce large amount of grass carp growth hormone (gcGH) of biological activity for further use,this study aimed at high yield expression of gcGH in the yeast Pichia pastoris .The gcGH cDNA was inserted into a yeast vector,pGAPZ α B.Under the control of the promoter GAP (glyceraldehyde 3 phosphate dehydrogenase),a 22 kilodalton protein,similar to the molecular weight of native growth hormone,was detected in the supernatant of transformed yeast.It led to a level as high as of 50 mg/L.This protein could be specifically immunoprecipitated with rabbit polyclonal antibodies raised against gcGH.A receptor ELISA Sandwich method was also perfomed to verify the biological activity of the in vitro expressed gcGH.

关 键 词：草鱼生长激素 毕赤酵母 表达 ELISA 

分 类 号：Q953[生物学—动物学]