摘要
目的建立一种以细菌染色体上1个已知拷贝管家基因作为基准、通过荧光定量PCR 检测其他被检基因拷贝数的方法。方法利用荧光定量PCR检测El Tor型霍乱弧菌染色体上目的基因zot与基准基因thyA的Ct(threshold cycle)值差值,根据已知拷贝数菌株N16961的两基因Ct值之差来推算待测菌株中zot基因的拷贝数。结果利用thyA基因作参照,确定11株El Tor型霍乱弧菌中 zot基因的拷贝数在1-5个之间,对较少拷贝数的检测与Southern blot确认的相同,对多拷贝数检测优于Southern blot的判定。结论本方法可以准确检测菌株染色体上的基因拷贝数。
Objective To establish a real time quantitative PCR method for detecting gene copy numbers in chromosome DNA of bacteria. Methods Ct values of genes zot and thyA of Vibrio cholerae strains are detected, and the difference of Ct values between these two genes is identified. The copy number of gene zot of one strain is determined with the reference of Ct value difference of strain N16961. Results Copy numbers of zot gene among 11 of detected Vibr/o cho/erae E1 Tot strains were determined from 1 to 5 with thyA as the reference. It showed that the established method is as good as ,Southern blot in terms of detecting low copy numbers of genes, and is better for determining high copy numbers. Conclusion The quantitative PCR can be applied to accurately detect copy numbers of genes in chromosome DNA of bacteria strains.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第4期379-382,共4页
Chinese Journal of Microbiology and Immunology
基金
国家十五科技攻关计划课题资助(2003BA712A05-04)国家重大基础研究项目(973)课题(G1999054102)
