基于PCR方法的基因序列全长获取策略

中国生物工程杂志

2012, Vol. 32

Issue (11): 115-123

综述

基于PCR方法的基因序列全长获取策略

李昆鹏^1,2, 朱化彬², 郝海生², 赵学明², 冯荣³, 秦彤², 张林波¹, 王栋²

1. 吉林农业大学动物科技学院长春 130118;
2. 中国农业科学院北京畜牧兽医研究所农业部畜禽遗传资源与利用重点开放实验室北京 100193;
3. 乌兰察布职业学院集宁 012000

The Strategies of Obtaining Full-length Sequence by PCR Amplification Technology

LI Kun-peng^1,2, ZHU Hua-bin², HAO Hai-sheng², ZHAO Xue-ming², FENG Rong³, QIN Tong², ZHANG Lin-bo¹, WANG Dong²

1. College of Animal Science ＆Technology, Jilin Agriculture University, Changchun 130118, China;
2. The Key Laboratory for Farm Animal Genetic Resources and Utilization of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China;
3. College of Wulanchabu Vocational, Jining 012000, China

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摘要： 基因全长序列是研究新基因功能的前提基础,基于PCR(polymerase chain reaction)建立了反向PCR、外源接头介导PCR和随机引物PCR三类基因序列全长扩增方法。虽然早期建立的反向PCR连接效率较低,易产生较高的非特异性扩增,但简单快捷的特点使其在鉴定T-DNA插入位点、基因融合位点等方面应用较多;外源接头介导PCR则因特异性的提高而备受欢迎,并衍生出单特异引物PCR、捕获PCR和步降PCR等多种技术,取得了较理想的扩增效果;而在大量样品分析中,自动化程度较高的随机引物PCR则更具优势。随着功能强大的聚合酶、连接酶、反转录酶的发现,推动了各种基于PCR的基因序列全长获取策略的发展,很多策略也因此而得以改进和整合,并获取更理想的实验结果,为研究基因结构、功能及其表达产物特性的研究提供了更完整的信息。

关键词： 全长序列; 染色体步移; RACE技术

Abstract: Full-length sequence is the basic premise of the study on genetic function of new gene, three amplification methods of the full-length sequence of gene, inverse PCR, adapter-mediated PCR and random primer PCR, have been established based on PCR(polymerase chain reaction). Although the early established inverse PCR is inefficient in the ligation procedure and bring forth more non-specific amplification, it's extensively used in the identification of T-DNA insertion sites and gene fusion sites for its simplicity and quickness. With the improvement of the specificity, adapter-mediated PCR is getting more and more popular, many novel technologies, such as single specific primer PCR, capture PCR and step-down PCR, were derived, and the ideal amplification effects were obtained. Recently, owing to its high degree of automation, random primer PCR is playing an important role in the analysis of a large number of samples. With the discovery of powerful enzymes, such as, DNA polymerase, ligase and reverse transcriptase, the development of the strategies for obtaining full-length sequence by PCR amplification technology were promoted, many strategies were improved and integrated, and better experimental results were obtained, it provided more complete information for the study of gene structure, function and transcript characteristics.

Key words: Full-length sequence Chromosome walking RACE technology

收稿日期: 2012-05-05 出版日期: 2012-11-25

ZTFLH:

Q781

基金资助: 国家"十二五"科技支撑计划资助项目(2011BAD19B02)

通讯作者: 张林波,电子信箱:cczlb@126.com;王栋,电子信箱:dwangcn2002@vip.sina.com.cn E-mail: cczlb@126.com;dwangcn2002@vip.sina.com.cn

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引用本文:

李昆鹏, 朱化彬, 郝海生, 赵学明, 冯荣, 秦彤, 张林波, 王栋. 基于PCR方法的基因序列全长获取策略[J]. 中国生物工程杂志, 2012, 32(11): 115-123.

LI Kun-peng, ZHU Hua-bin, HAO Hai-sheng, ZHAO Xue-ming, FENG Rong, QIN Tong, ZHANG Lin-bo, WANG Dong. The Strategies of Obtaining Full-length Sequence by PCR Amplification Technology. China Biotechnology, 2012, 32(11): 115-123.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I11/115

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[1]	李昆鹏, 朱化彬, 郝海生, 赵学明, 冯荣, 秦彤, 张林波, 王栋. 基于PCR方法的基因序列全长获取策略[J]. 中国生物工程杂志, 2012, 32(11): 115-123.
[2]	王萍, 江木兰, 张银波, 万霞, 梁焯, 龚阳敏. 载有磷酸甘油激酶基因启动子的新表达载体的构建以及在发酵性丝孢酵母中启动外源基因的表达研究[J]. 中国生物工程杂志, 2012, 32(03): 39-46.
[3]	李付鹏, 伍宝朵, 马朝芝, 傅廷栋. 基于PCR的染色体步移技术研究进展[J]. 中国生物工程杂志, 2010, 30(12): 87-94.

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