[1]杨运文,蒋伏欢,宋杰,等.重组工程法敲除恶臭假单胞菌KT2440的染色体基因[J].南京师大学报(自然科学版),2011,34(04):96-101.
 Yang Yunwen,Jiang Fuhuan,Song Jie,et al.Recombineering Mediated Chromosal Gene Knockout of Pseudomonas Putida KT2440[J].Journal of Nanjing Normal University(Natural Science Edition),2011,34(04):96-101.
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重组工程法敲除恶臭假单胞菌KT2440的染色体基因()
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《南京师大学报(自然科学版)》[ISSN:1001-4616/CN:32-1239/N]

卷:
第34卷
期数:
2011年04期
页码:
96-101
栏目:
生命科学
出版日期:
2011-12-20

文章信息/Info

Title:
Recombineering Mediated Chromosal Gene Knockout of Pseudomonas Putida KT2440
作者:
杨运文蒋伏欢宋杰尚广东
南京师范大学生命科学学院江苏省功能微生物与功能基因组学重点实验室,江苏南京210046
Author(s):
Yang YunwenJiang FuhuanSong JieShang Guangdong
Jiangsu Key Laboratory for Microbes and Functional Genomics,School of Life Sciences, Nanjing Normal University,Nanjing 210046,China
关键词:
恶臭假单胞菌KT2440基因敲除重组工程
Keywords:
Pseudomonas putida KT2440gene knockoutrecombineering
分类号:
Q78
摘要:
恶臭假单胞菌KT2440是在生物降解环境有机物和表达异源基因等方面具重要作用的环境微生物.基因敲除是研究基因和蛋白功能的必要手段.常规的基因敲除方法往往会烦琐耗时且易引入突变的基因克隆操作.本研究采用重组工程(即重组酶催化的PCR产物与基因组上同源片段之间的同源重组)来实现恶臭假单胞菌KT2440的基因敲除.本方法高效、简便、快捷且可以同时进行多个基因的操作,可成为通用的恶臭假单胞菌KT2440的遗传研究方法.
Abstract:
Pseudomonas putida KT2440 plays an important role in the degradation of environmental organic compounds and it is a good host for foreign gene expression. Gene knockout is essential to the investigations of genes and proteins. Classic chromosal gene knockout methods usually involve the gene cloning steps,which are often tedious,time consuming and tend to introduce mutations. In this paper,a recombineering strategy characterized by the homologous recombination between the PCR product and the genome,is used for the P. putida KT2440 gene knockout. This strategy is highly efficient,convenient,rapid and amendable for the simultaneously manipulation of many genes,and can be used as a general gene knockout method.

参考文献/References:

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相似文献/References:

[1]樊 丹,石牡丹,杨运文,等.基于染色体的重组工程系统的改进[J].南京师大学报(自然科学版),2013,36(01):99.
 Fan Dan,Shi Mudan,Yang Yunwen,et al.Improvement of the Chromosome-based Recombineering System[J].Journal of Nanjing Normal University(Natural Science Edition),2013,36(04):99.

备注/Memo

备注/Memo:
基金项目:“重大新药创制”科技重大专项课题( 2009ZX091032674) .通讯联系人: 尚广东,副教授,研究方向: 微生物药物的研发. E-mail: shanggd@ hotmail. Com
更新日期/Last Update: 2013-03-21