中国农学通报
中國農學通報
중국농학통보
CHINESE AGRICULTURAL SCIENCE BULLETIN
2013年
17期
70-74
,共5页
江小斌%黄海宁%杨小强%黄晓南%刘年锋%张善霹%张浩
江小斌%黃海寧%楊小彊%黃曉南%劉年鋒%張善霹%張浩
강소빈%황해저%양소강%황효남%류년봉%장선벽%장호
大黄鱼%生长激素%胞外表达%Pichia pastoris
大黃魚%生長激素%胞外錶達%Pichia pastoris
대황어%생장격소%포외표체%Pichia pastoris
Pseudosciaena crocea%Growth Hormone%Extracellular Expression%Pichia pastoris
为大量获得具有生物活性的大黄鱼生长激素(Pseucdosciaena crocea,Growth Hormone,pcGH),利用本实验室分离的天然大黄鱼生长激素基因在毕赤酵母表达系统中的表达进行了研究。把pcGH克隆到酵母整合型质粒载体中构建重组表达载体 pPIC9K-pcGH,电击转化将pcGH 转化进his4缺陷型毕赤酵母GSll5菌株染色体上,经甲醇诱导得以高效表达,SDS-PAGE证实了表达产物为大黄鱼生长激素,表达量约为62.5 mg/L。
為大量穫得具有生物活性的大黃魚生長激素(Pseucdosciaena crocea,Growth Hormone,pcGH),利用本實驗室分離的天然大黃魚生長激素基因在畢赤酵母錶達繫統中的錶達進行瞭研究。把pcGH剋隆到酵母整閤型質粒載體中構建重組錶達載體 pPIC9K-pcGH,電擊轉化將pcGH 轉化進his4缺陷型畢赤酵母GSll5菌株染色體上,經甲醇誘導得以高效錶達,SDS-PAGE證實瞭錶達產物為大黃魚生長激素,錶達量約為62.5 mg/L。
위대량획득구유생물활성적대황어생장격소(Pseucdosciaena crocea,Growth Hormone,pcGH),이용본실험실분리적천연대황어생장격소기인재필적효모표체계통중적표체진행료연구。파pcGH극륭도효모정합형질립재체중구건중조표체재체 pPIC9K-pcGH,전격전화장pcGH 전화진his4결함형필적효모GSll5균주염색체상,경갑순유도득이고효표체,SDS-PAGE증실료표체산물위대황어생장격소,표체량약위62.5 mg/L。
This study aimed at high-yield expression of the pcGH gene which was separated from natural large yellow croaker in the yeast Pichia pastoris in order to produce amount of large yellow croaker growth hormone (Pseucdosciaena crocea, Growth Hormone, pcGH) with the feature of biological activity for further use. The pcGH gene was first cloned into the integrative expression vector of methylotrophic yeast P. Pastoris to form the recombinant vector pPIC9K-pcGH, and the linearized recombinant plasmid was then transformed into the chromosome of the HIS4 mutant yeast GS115 by electroporation, and it could finally get high-yield expression induced by methanol. SDS-PAGE confirmed that the expression product was pcGH, and it led to a level as high as of 62.5 mg/L.