| 海洋球石藻病毒(Coccolithovirus)硫氧还蛋白(Trx)在毕赤酵母(Pichia pastoris)中的表达及其活性分析 |
| |
| 引用本文： | 蔡艺钦,张稚兰,罗邦彬,刘静雯.海洋球石藻病毒(Coccolithovirus)硫氧还蛋白(Trx)在毕赤酵母(Pichia pastoris)中的表达及其活性分析[J].海洋与湖沼,2012,43(5):905-910. |
| |
| 作者姓名： | 蔡艺钦 张稚兰 罗邦彬 刘静雯 |
| |
| 作者单位： | 集美大学生物工程学院,厦门,361021 |
| |
| 基金项目： | 福建省自然科学基金项目, 2010J01261 号; 集美大学中青年创新团队专项基金资助, 2010A007 号 |
| |
| 摘 要： | 从实验室保存的pBS-Trx重组质粒中克隆球石藻病毒EhV-Trx基因,构建毕赤酵母重组表达载体pPIC9K-EhV-Trx,将重组质粒电转化毕赤酵母GS115,诱导分泌表达并对重组蛋白进行二硫键还原酶活性分析。结果表明,EhV-Trx基因开放阅读框为591bp,编码197个氨基酸;在毕赤酵母GS115中成功诱导表达重组EhV-Trx,经SDS-PAGE分析目的蛋白分子量约为27.8kDa;重组EhV-Trx具有二硫键还原酶的活性,能有效打开胰岛素A、B两条链的二硫键,有望开发成一种新型的硫氧还蛋白脱敏制剂应用于食品安全领域。
|
| 关 键 词： | 海洋球石藻病毒(EhV) 硫氧还蛋白 毕赤酵母 分泌表达 活性分析 |
| 收稿时间： | 2012/4/10 0:00:00 |
| 修稿时间： | 2012/6/15 0:00:00 |
EXPRESSION AND ACTIVITY ANALYSIS OF THIOREDOXIN (Trx) FROM MARINE COCCOLITHOPHORID EMILIANIA HUXLEYI VIRUS IN PICHIA PASTORIS |
| |
| CAI Yi-Qin,ZHANG Zhi-Lan,LUO Bang-Bin and LIU Jing-Wen.EXPRESSION AND ACTIVITY ANALYSIS OF THIOREDOXIN (Trx) FROM MARINE COCCOLITHOPHORID EMILIANIA HUXLEYI VIRUS IN PICHIA PASTORIS[J].Oceanologia Et Limnologia Sinica,2012,43(5):905-910. |
| |
| Authors: | CAI Yi-Qin ZHANG Zhi-Lan LUO Bang-Bin and LIU Jing-Wen |
| |
| Institution: | College of Bio-Engineering, Jimei University;College of Bio-Engineering, Jimei University;College of Bio-Engineering, Jimei University;College of Bio-Engineering, Jimei University |
| |
| Abstract: | Thioredoxin (Trx), a small molecule ubiquitous multifunctional acidic protein, was found to reduce disulfide bonds of protein (convert SOS to 2 SH) and thereby mitigate the allergenicity of food. Emiliania huxleyi virus (EhV) is the first virus containing Trx gene reported. Here the gene of EhV-Trx was amplified from the recombinant plasid of pBS-EhV99B1-Trx by PCR. Ehv-Trx gene was inserted yeast expression vector pPIC9K to construct recombinant expression plasmid pPIC9K-EhV99B1-Trx. After sequencing, the recombinant expression plasmid was transformed into Pichia pastoris GS115 by electroporation method. The recombinant protein EhV99B1-Trx reduction activity was detected. The results showed that: the open-reading frame (ORF) of EhV99B1-Trx encoded a protein of 197 amino acids; the recombinant EhV99B1-Trx was successfully induced expression in P. pastoris GS115 and the target protein molecular mass was about 27.8kDa; as found for other proteins with intramolecular disulfide bonds, insulin were reduced specifically by the recombinant EhV99B1-Trx and the treated product was relatively stable, which indicated that the EhV99B1-Trx, as a new kind of thioredoxin, do have the potential in food safety areas. |
| |
| Keywords: | Emiliania huxleyi virus (EhV) Thioredoxin (Trx) Pichia pastoris Secreted expression Activity analysis |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《海洋与湖沼》浏览原始摘要信息 |
| 点击此处可从《海洋与湖沼》下载免费的PDF全文 |
|
