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| 脊尾白虾热休克蛋白HSP90基因的原核表达与鉴定 | |
| 作者姓名： | 韩俊英 李健 李吉涛 陈萍 李华 |
| 作者单位： | 1. 大连海洋大学,116023;农业部海洋渔业资源可持续利用重点开放实验室中国水产科学研究院黄海水产研究所,青岛266071 2. 农业部海洋渔业资源可持续利用重点开放实验室中国水产科学研究院黄海水产研究所,青岛,266071 3. 大连海洋大学,116023 |
| 基金项目： | 公益性行业(农业)科研专项(nyhyzx07-042,200803012); 国家虾产业技术体系(CARS-47); 科技部农业科技成果转化资金项目(2010GB23260589)共同资助 |
| 摘 要： | 将脊尾白虾热休克蛋白90基因克隆到原核表达载体pET-30a中,经酶切验证和DNA测序鉴定后,将重组质粒转化表达宿主大肠杆菌Rosetta,优化温度、时间、IPTG和OD600表达条件进行诱导表达,收集菌液,进行SDS-PAGE和质谱检测,并用Quantity one软件分析蛋白表达水平。结果表明,成功构建了含脊尾白虾HSP90基因的重组表达载体pET-30a-HSP90,表达目的蛋白相对分子量为82.7kD,为HSP90蛋白。通过条件优化认为重组菌株Rosetta/pET-30a-HSP90的最佳诱导温度为37℃,最佳IPTG浓度为1.0mmol/L,最佳诱导时机和诱导时间分别为0.58h、7h。 |
| 关 键 词： | 脊尾白虾 热休克蛋白90 原核表达 |
| 收稿时间： | 2010/12/16 0:00:00 |
| 修稿时间： | 2011/3/31 0:00:00 |
High level prokaryotic expression and identification of heat shock protein 90 in Exopalaemon carinicauda |
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| Authors: | HAN Jun-ying LI Jian LI Ji-tao CHEN Ping LI Hua |
| Institution: | HAN Jun-ying 1,2 LI Jian2 LI Ji-tao2 CHEN Ping2 LI Hua1 (1Dalian Ocean University,116023) (2Key Laboratory for Sustainable Utilization of Marine Fisheries Resources,Ministry of Agriculture,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071) |
| Abstract: | The aim of this study was to set up a high level prokaryotic expression of heat shock protein 90(HSP90) of Exopalaemon carinicauda in E. coli Rosetta. The HSP90 gene of E. carinicauda was cloned into prokaryotic expression vector pET-30a, which was confirmed by double-endonuclease digestion and DNA sequencing. The recombinant vector was transformed into E. coli Rosetta and was induced to express under different temperatures, durations, IPTG concentrations and OD600. The expressed product was identified by S... |
| Keywords: | Exopalaemon carinicauda Heat shock protein 90 (HSP90) Prokaryotic expression |
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