| HSV_1-TK基因原核表达载体的构建及表达检测 |
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| 引用本文： | 王丽群,孟祥晨.HSV_1-TK基因原核表达载体的构建及表达检测[J].东北农业大学学报,2009,40(2). |
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| 作者姓名： | 王丽群 孟祥晨 |
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| 作者单位： | 乳品科学教育部重点实验室,东北农业大学食品学院,哈尔滨,150030 |
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| 基金项目： | 东北农业大学博士基金 |
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| 摘 要： | 探讨自杀基因HSV1-TK原核表达情况,为进一步将细菌载体应用于肿瘤基因治疗奠定基础。首先以质粒pHSV-106为模板,通过PCR获得HSV1-TK基因克隆并将其与原核表达载体pGEX-6P-1连接,重组质粒经鉴定后将其转化到E.coli BL21,观察其蛋白表达情况。结果表明,TK基因全长1 128 bp,与理论值相符。经测序发现插入到载体pGEX-6P-1上的DNA片段与TK基因的同源性为100%;SDS-PAGE可明显看出大小为41 ku的TK酶表达,并随时间的延长表达量不断增加。试验证明来源于单纯疱疹病毒的胸苷激酶能够实现原核表达,这一结果是应用细菌载体携带自杀基因TK治疗肿瘤的前提。
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| 关 键 词： | HSV1-TK基因 原核表达 基因治疗 肿瘤 |
Construction of prokaryotic expression vector for HSV_1-TK and detection of expressed protein |
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| WANG Liqun,MENG Xiangchen.Construction of prokaryotic expression vector for HSV_1-TK and detection of expressed protein[J].Journal of Northeast Agricultural University,2009,40(2). |
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| Authors: | WANG Liqun MENG Xiangchen |
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| Institution: | WANG Liqun,MENG Xiangchen(Key Laboratory of Dairy Science,Ministry of Education,Food College,Northeast Agricultural University,Harbin 150030,China) |
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| Abstract: | This paper discussed the self-destruction gene HSV1-TK prokaryotic expression in order to establish the foundation of bacteria vector applied in the tumor gene therapy.Firstly,plasmid pHSV-106 was taken as the template,the clone of HSV1-TK gene was acquired through PCR and connected to the prokaryotic expression vector pGEX-6P-1,then recombinant plasmid was identified and transformed into E.coli BL21(DE3) competence cell to observe the protein expression.The result showed that the length of TK gene was 1 12... |
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| Keywords: | HSV1-TK gene prokaryotic expression gene therapy tumor |
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