日本鳗鲡黄体生成素在毕氏酵母中的表达  

Expression of Recombinant Follicle Stimulating Hormone of Japanese Eel Anguilla japonica Using Pichia pastoris

作　　者：黄名锡 戴荣贵 陈磊 王梦洋[1,2,3] 谢仰杰 陈天圣 HUANG Mingxi;DAI Ronggui;CHEN Lei;WANG Mengyang;XIE Yangjie;CHEN Tiansheng(State Key Laboratory of Mariculture Breeding,Xiamen 361021,China;Fisheries College,Jimei University,Xiamen 361021,China;Engineering Research Center of the Modern Technology for Eel Industry,Ministry of Education,Xiamen 361021,China)

机构地区：[1]海水养殖生物育种全国重点实验室,福建厦门361021 [2]集美大学水产学院,福建厦门361021 [3]鳗鲡现代产业技术教育部工程研究中心,福建厦门361021

出　　处：《集美大学学报(自然科学版)》2025年第1期1-8,共8页Journal of Jimei University:Natural Science

基　　金：福建省自然科学基金项目“海水驯养对鳗鲡卵巢发育的影响及分子机理”(2023J01145);福建省自然科学基金项目“鳗鲡促卵泡生产激素FSH的表达和功能分析”(2024J01104);集美大学科学研究基金项目“基因编辑在青鳉分子抗病育种的研究”(ZQ2020003)。

摘　　要：日本鳗鲡(Anguilla japonica)的人工繁育技术尚未解决,其重要原因是饲养条件下鳗鲡性腺不能发育成熟。黄体生成素(luteinizing hormone,LH)是促性腺激素的主要成分。为了解黄体生成素的功能,利用毕氏酵母生产具有生物活性的重组日本鳗鲡黄体生成素,构建了LHβ和GTHα亚基cDNA串联的669 bp的真核表达载体,通过转化X-33酵母菌株,成功表达了日本鳗鲡黄体生成素重组蛋白。重组蛋白糖基化后相对分子质量为45~75 ku,经过N-糖苷酶(glycopeptidase F)去糖基化后蛋白呈现单一条带,且相对分子质量与理论大小相符。在0.5%甲醇诱导下,96 h重组蛋白表达量最高,纯化后可获得3~4 mg/L的蛋白量。A key hurdle is the failure of eel gonads to mature under artificial feeding conditions,linked closely to the production and secretion of gonadotropins.Among these,luteinizing hormone(LH)plays a pivotal role.To unravel the role of Japanese eel LH,biologically active recombinant LH was synthesized using Pichia pastoris.The recombinant Japanese eel LH was successfully expressed by transforming the X-33 yeast strain with a 669 bp expression vector containing LHβand GTHαsubunit cDNA.The protein exhibited a molecular weight ranging from 45 ku to 75 ku.Following deglycosylation with glycopeptidase F enzyme,the protein displayed a single band around 25 ku,aligning with the anticipated size.Optimal expression,peaking at 96 hours with 0.5%methanol induction,yielded purified protein at concentrations of 3-4 mg/L.

关 键 词：日本鳗鲡 黄体生成素 重组蛋白 酵母表达 

分 类 号：S961[农业科学—水产养殖]