| 水生栖热菌海藻糖合成酶基因的克隆及真核表达 |
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| 引用本文： | 刘俊梅,聂海彦,郑微微,胡耀辉.水生栖热菌海藻糖合成酶基因的克隆及真核表达[J].食品科学,2010,31(23):267-270. |
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| 作者姓名： | 刘俊梅 聂海彦 郑微微 胡耀辉 |
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| 作者单位： | 吉林农业大学食品科学与工程学院 |
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| 基金项目： | 国家科技部成果转化基金项目(2007GB2B10072 )；分子酶工程教育部重点实验室平台基地建设项目(20100419) |
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| 摘 要： | 通过基因工程技术合成海藻糖合酶Tres 全基因，插入到毕赤酵母菌表达载体pPICZα中。电转化毕赤酵母菌GS115 后，经Zeocin 抗性筛选阳性菌株。用PCR 和全基因测序鉴定目的基因的表达。通过诱导表达目的蛋白，并以SDS-PAGE 和Western blotting 进行鉴定。成功地构建了pPICZα-Tres 重组质粒，并证明目的基因已整合于毕赤酵母菌的基因组。
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| 关 键 词： | 海藻糖合酶 Tres 全基因 基因克隆 真核表达 |
| 收稿时间： | 2010-09-14 |
Cloning and Eukaryotic Expression of Trehalose Synthase Gene from Thermus aquaticus FL-03 |
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| LIU Jun-mei,NIE Hai-yan,ZHENG Wei-wei,HU Yao-hui.Cloning and Eukaryotic Expression of Trehalose Synthase Gene from Thermus aquaticus FL-03[J].Food Science,2010,31(23):267-270. |
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| Authors: | LIU Jun-mei NIE Hai-yan ZHENG Wei-wei HU Yao-hui |
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| Affiliation: | (College of Food Science and Engineering, Jilin Agricultural University, Changchun 130118, China) |
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| Abstract: | The full-length trehalose synthase gene (Tres) was synthesized by genetic engineering and inserted into the Pichia expression vector pPICZα. The expression vector carrying Tres gene was transformed into the competent cells of Pichia GS115 by means of electroporation, and Zeocin resistance screening was then performed to find positive strains. PCR and gene sequencing were used to identify the expression of the target gene. The expressed protein was analyzed by SDS-PAGE and Western blotting. The results suggested that the recombinant plamid pPICZα-Tres was successfully constructed and that the target gene had been integrated into the Pichia genome. |
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| Keywords: | trehalose synthase Tres gene gene cloning eukaryotic expression |
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