Expression of A Recombinant Luteinizing Hormone of Marble Eels，Anguilla marmorata Using Methylotropic Yeast，Pichia pastoris

MAO Limin; ZHANG Yong; LI Yangyuan; HUANG Hai; LI Bo; LI Wensheng; CHEN Guohua; LIN Haoran; 

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Expression of A Recombinant Luteinizing Hormone of Marble Eels，Anguilla marmorata Using Methylotropic Yeast，Pichia pastoris

更新时间：2023-12-11

- Expression of A Recombinant Luteinizing Hormone of Marble Eels，Anguilla marmorata Using Methylotropic Yeast，Pichia pastoris
- Acta Scientiarum Naturalium Universitatis SunYatseni Vol. 51, Issue 2, Pages: 86-90(2012)
- 作者机构：
  
  1. ．海南大学海洋学院暨热带生物资源教育部重点实验室,海南,海口,570228
  2. 2．中山大学有害生物控制与资源利用国家重点实验室∥中山大学水生经济动物研究所暨广东省水生经济动物良种繁育重点实验室,广东,广州,510275
- 作者简介：
- 基金信息：
- DOI：
  CLC：
- Published：2012，
  
  Published Online：25 March 2012，
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MAO Limin, ZHANG Yong, LI Yangyuan, et al. Expression of A Recombinant Luteinizing Hormone of Marble Eels，Anguilla marmorata Using Methylotropic Yeast，Pichia pastoris. [J]. Acta Scientiarum Naturalium Universitatis SunYatseni 51(2):86-90(2012)
DOI：

MAO Limin, ZHANG Yong, LI Yangyuan, et al. Expression of A Recombinant Luteinizing Hormone of Marble Eels，Anguilla marmorata Using Methylotropic Yeast，Pichia pastoris. [J]. Acta Scientiarum Naturalium Universitatis SunYatseni 51(2):86-90(2012) DOI：

摘要

采用聚合酶链反应从花鳗鲡垂体cDNA中分别扩增出

GTHα、LHβ

基因，再利用搭桥技术，中间以linker将两个基因连接成单个基因

LHβα

。然后利用双酶切、连接，将

LHβα

基因连接到载体pPICZαA中，构建出促黄体生成素的酵母共表达载体pPICZαA-

LHβα

。再利用电转化法将其转入到酵母野生型菌株X-33中，在含不同浓度的zeocinYPDS平板上筛选高拷贝转化子，阳性克隆经φ=0.7%甲醇诱导表达，表达产物采用SDS-PAGE和western blot进行分析，得到所表达的产物相对分子质量约为45 000。

Abstract

The

GTHα、LHβ

gene fragments were amplified by PCR

which the template was pituitary cDNA of marble eels. In the way of bridging，the two fragments were connected to a single gene

LHβα



a linker inserted between them. The 

LHβα

gene fragment was subcloned to pPICZαA with double enzyme digestion and connection

constructing eukaryotic coexpression plasmid pPICZα-

LHβα

. The recombinant plasmid was transfected into a native yeast strain X-33 by electrotransformation. The high-copy clones were selected by the YPDS plates containing different concentrations of Zeocin. The positive clones were induced by 0.7% methanol and expression products were tested by SDS-PAGE and western blot

and its molecular weight was determined as about 45 000.

关键词

花鳗鲡促黄体生成激素linker共表达载体pPICZαA-LHβα毕赤酵母

Keywords

Anguilla marmorataluteinizing hormonelinkerco-expression plasmid pPICZαA-LHβαPichia pastoris

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Related Institution

State Key Laboratory of Biocontrol, The Key Laboratory of Gene Engineering of Ministry of Education and Biotechnology Research Center, Sun Yatsen University

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School of Life Sciences,Sun Yatsen University

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