| 美洲大蠊3-磷酸甘油醛脱氢酶基因的克隆及表达 |
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| 引用本文： | 龙高群,王赟,张春林.美洲大蠊3-磷酸甘油醛脱氢酶基因的克隆及表达[J].生物技术通报,2012(4):113-117. |
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| 作者姓名： | 龙高群 王赟 张春林 |
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| 作者单位： | 1. 贵阳医学院生物学教研室,贵阳,550004
2. 贵阳医学院生物技术教研室,贵阳,550004 |
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| 基金项目： | 贵州省社发攻关项目，贵阳医学院研究生创新计划专项基金 |
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| 摘 要： | 旨在通过5’-RACE获得美洲大蠊3-磷酸甘油醛脱氢酶(GAPDH)基因的全长cDNA序列,进行生物信息学分析,构建原核表达载体,诱导重组蛋白表达,为进一步研究其功能奠定基础.通过3’-RACE技术,PCR扩增获取编码美洲大蠊GAPDH蛋白的全长cDNA序列；采用生物信息学方法推导出该序列编码的氨基酸序列及其理化性质；预测信号肽、蛋白疏水性、可溶性、跨膜区结构、二级结构、三级结构,并构建系统发育树；构建原核表达载体pET28a-GAPDH,IPTG诱导重组蛋白表达,并用Histag抗体Western blotting验证.结果显示,美洲大蠊GAPDH基因,其完整阅读框含999个碱基,编码332个氨基酸.序列分析显示,该蛋白与家蚕GAPDH相似性为89％,具有GAPDH保守功能域,经IPTG诱导获得重组蛋白.
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| 关 键 词： | 美洲大蠊 3-磷酸甘油醛脱氢酶 克隆 序列分析 原核表达 |
Molecular Cloning and Expression of Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) Gene of Periplaneta americana |
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| Long Gaoqun
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Wang Yun
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Zhang Chunlin.Molecular Cloning and Expression of Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) Gene of Periplaneta americana[J].Biotechnology Bulletin,2012(4):113-117. |
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| Authors: | Long Gaoqun Wang Yun Zhang Chunlin |
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| Institution: | 1(1Department of Biology,Guiyang Medical College,Guiyang 550004;2Department of Biotechnology,Guiyang Medical College,Guiyang550004) |
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| Abstract: | It was to clone the GAPDH gene of Periplaneta american,analyze its sequence,express recombinant protein of thePeripla-neta americanaGAPDH,predict the protein structure.Base on a fragment sequence of the GAPDH fromPeriplaneta americancDNA library,obtained full-length GAPDH sequence by RACE.The recombinant plasmid pET28a(+)-GAPDH was built to express His-GAPDH fusion protein in E.coli BL21(DE3)by IPTG induction,and identified by SDS-PAGE and Western blotting.Results showed the Periplaneta american GAPDH gene include an reading fragment of 999 bp code for 332 amino acid residues.It revealed 89% identity with GAPDH gene ofBombyx mori. |
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| Keywords: | Periplaneta americana GAPDH Clone Sequence analysis Prokaryotic express |
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