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曼氏无针乌贼(Sepiella maindroni)sigma-型谷胱甘肽硫转移酶基因的克隆及重组表达
余作奔, 王春琳, 母昌考, 宋微微, 李荣华
宁波大学海洋学院
摘要:
采用EST结合RACE技术进行了曼氏无针乌贼(Sepiella maindroni)sigma-型GST(SmGST)基因的克隆。结果表明, 该基因的cDNA全长为838bp, 其中5′非编码区为73bp, 3′非编码区为150bp, 开放阅读框为615bp, 编码的蛋白含有204个氨基酸。该基因的氨基酸序列中包含1个典型的GST-N结构域和一个GST-C结构域, 与栉孔扇贝和长牡蛎的sigma-型GST的相似度分别为40.3%和39.3%。将该基因的编码区重组到pET-21(a+)载体后在大肠杆菌中得到诱导表达。重组SmGST的GST活力为(12.22± 0.92)U/mg蛋白。
关键词:  曼氏无针乌贼  谷胱甘肽硫转移酶  分子克隆  重组表达
DOI：10.11693/hyhz201302035035
分类号:
基金项目:国家自然科学基金项目资助, 41176124 号, 41206114 号; 教育部博士点基金资助, 20103305110003 号; 浙江省自然科学基金重点项目资助, Z3110482 号; 浙江省自然科学基金项目资助, LQ12C19002 号。
MOLECULAR CLONING AND RECOMBINANT EXPRESSION OF A SIGMA-LIKE GLUTATHIONE S-TRANSFERASE FROM SEPIELLA MAINDRONI
YU Zuo-Ben, WANG Chun-Lin, MU Chang-Kao, SONG Wei-Wei, LI Rong-Hua
School of Marine Science of Ningbo University
Abstract:
The sigma-like glutathione S-transferase in Sepiella maindroni was cloned by EST and RACE method. The full sequence was of 838bp, containing an open reading frame (ORF) of 615bp, that encoded 204 amino acid residues and a 3′untranslated region of 150bp and a 5′UTR of 73bp. The deduced amino acid sequence of this gene showed 39.3%— 40.3% identity to the sequences of the sigma-class GSTs from Chlamys farreri and Crassostrea gigas. This gene was expressed in Escherichia coli with the pET-21(a+) vector. The GST activity of the recombinant SmGST was (12.22±0.92)U/mg protein.
Key words:  Sepiella maindroni  glutathione S-transferase  molecular cloning  recombinant expression
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