[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):99-103.
　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(01):99-103.
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基于染色体的重组工程系统的改进()
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《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

卷:
第36卷
期数:
2013年01期
页码:
99-103
栏目:
生命科学
出版日期:
2013-03-31

文章信息/Info

Title:
Improvement of the Chromosome-based Recombineering System
作者:
樊 丹; 石牡丹; 杨运文; 尚广东
南京师范大学生命科学学院,江苏省微生物工程技术研究中心,江苏省微生物与功能基因组学重点实验室,江苏 南京 210023
Author(s):
Fan Dan; Shi Mudan; Yang Yunwen; Shang Guangdong
Jiangsu Engineering and Technology Research Center for Microbiology,Jiangsu Key Laboratory for Microbes and Functional Genomics,School of Life Sciences,Nanjing Normal University,Nanjing 210023,China
关键词:
重组工程; 基因敲除; 重组效率; 单链寡核苷酸修复; 双链断裂修复
Keywords:
recombineering; knock-out; recombination efficiency; SSOR; DSBR
分类号:
Q819
摘要:
重组工程是指通过重组酶催化DNA之间的同源重组,实现DNA克隆和修饰的一种基因工程技术.本研究从重组酶基因整合至大肠杆菌DH10B所获得的基于染色体的重组工程系统菌株LS-GR出发,利用菌株自身的重组工程系统和双链断裂修复功能,敲除了编码5’->3’单链DNA外切酶的recJ基因和编码3’->5’ DNA外切核酸酶的sbcB基因,获得重组效率得以提高的菌株LS2002和LS2004.单链寡核苷酸修复和双链断裂修复实验表明所得菌株较LS-GR的重组效率有明显提高,对需要重组效率高的重组工程实验具有重要的应用价值.
Abstract:
Recombineering is a genetic engineering technology for the DNA cloning and modification based on the recombinase-catalyzed homologous recombination.In this study,starting from previous obtained LS-GR,a chromosome-based recombineering strain,the 5’->3’ single stranded DNA exonuclease coding gene recJ and exonucleolytic cleavage in the 3’->5’ DNA exonuclease coding gene sbcB were knocked out via LS-GR’s recombineering function as well as its inherent double strand break repair(DSBR)function.Compared with LS-GR,the obtained strains LS2002 and LS2004 show higher single strand oligonucleotide repair(SSOR)activity and DSBR activities,showing that they have the potential to be used in the recombineering experiments for high recombination efficiency.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2012-07-03.
基金项目:江苏省科技厅工程中心创新能力提升项目、南京市科委工程中心创新能力提升项目(201105058).
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: 2013-03-31