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| 限制性显示-聚合酶链反应扩增苏云金杆菌基因片段的克隆与分析 | |
| 引用本文： | 李侠,马文丽,庞义,张宝,姜立,郑文岭. 限制性显示-聚合酶链反应扩增苏云金杆菌基因片段的克隆与分析[J]. 南方医科大学学报, 2003, 23(4): 323-325 |
| 作者姓名： | 李侠 马文丽 庞义 张宝 姜立 郑文岭 |
| 作者单位： | 1. 第一军医大学分子生物学研究所,广东,广州,510515 2. 中山大学生命科学学院,广东,广州,510632 3. 广州军区广州总医院分子肿瘤学研究所,广东,广州,510010 |
| 基金项目： | 国家自然科学基金(39880032)，广州市重点科技攻关项目(99-Z-022-01)~~ |
| 摘 要： | 目的运用限制性显示-PCR技术(RD-PCR)扩增苏云金杆菌(Bacillusthuringiensis,Bt)基因片段,并进行克隆、测序分析。方法设计特异性引物扩增长片段Bt基因,对扩增产物进行RD-PCR扩增,扩增后的产物克隆至pMD18-T载体并进行快速鉴定。提取阳性克隆质粒进行测序分析。结果运用RD-PCR技术,将长片断的基因(2 000-3 000 bp)分为多个短的片段,片段长度较为均一(200~600 bp)。测序结果表明,所扩增的片段均属于特异扩增的Bt基因。结论运用RD-PCR技术可以将长片段基因进行片段化,使其适用于基因芯片的探针。 |
| 关 键 词： | 限制性显示-聚合酶链反应 苏云金杆菌 基因克隆 |
| 文章编号： | 1000-2588(2003)04-0323-03 |
| 修稿时间： | 2002-11-10 |
Cloning and sequence analysis of Bacillus thuringiensis gene fragments isolated by restriction digest PCR |
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| LI Xia ,MA Wen-li ,PANG Yi ,ZHANG Bao ,JIANG Li ,ZHENG Wen-Ling Institute of Molecular Biochemistry,First Military Medical University,Guangzhou ,China, College of Life Science,Sun Yet-sen University,Guangzhou ,China. Cloning and sequence analysis of Bacillus thuringiensis gene fragments isolated by restriction digest PCR[J]. Journal of Southern Medical University, 2003, 23(4): 323-325 | |
| Authors: | LI Xia MA Wen-li PANG Yi ZHANG Bao JIANG Li ZHENG Wen-Ling Institute of Molecular Biochemistry First Military Medical University Guangzhou China College of Life Science Sun Yet-sen University Guangzhou China |
| Affiliation: | LI Xia 1,MA Wen-li 1,PANG Yi 2,ZHANG Bao 1,JIANG Li 1,ZHENG Wen-Ling 31 Institute of Molecular Biochemistry,First Military Medical University,Guangzhou 510515,China, 2 College of Life Science,Sun Yet-sen University,Guangzhou 510632,China, 3 Institute of Molecular Oncology,Guangzhou General Hospital of Guangzhou Command,Guangzhou 510010,China |
| Abstract: | Objective To clone and analyze Bacillus thuringiensis gene fragments isolated by restriction digest PCR (RD-PCR). Method Specific primers were designed to amplify the genes of Bacillus thuringiensis israelensis(Bti), and the PCR products were classified and re-amplified by RD-PCR to obtain the fragments for subsequent purification and cloning into the pMD18-T vectors, followed by rapid identification. The recombinant plasmids were extracted from positive clones and the target gene fragments were sequenced. Results Sequence analysis showed that all the fragments amplified were Bti genes. Conclusion RD-PCR is reliable in breaking down large gene fragments into confined and shorter gene fragments for preparing microarray probes. |
| Keywords: | restriction digest-PCR Bacillus thuringiensis gene clone |
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