摘要
金黄色葡萄球菌存在两个核酸酶编码基因,一个是葡萄球菌核酸酶(Staphylococcal nuclease,SNase),命名为nuc1,另一个是耐热核酸酶(Thermonuclease,TNase),命名为nuc2,nuc2是一个新的候选基因,以往认为金黄色葡萄球菌中的核酸酶只源于一个编码基因nuc1,为了进一步研究nuc2基因的功能,首先要将金黄色葡萄球菌nuc1基因缺失。研究目的就是通过构建同源重组质粒pBT2Δnuc1,将其电转入金黄色葡萄球菌菌株RN4220中,获得nuc1基因缺失突变株。经过了7轮培养和筛选,同源重组几率为2%(7/345),筛选出的nuc1突变株用PCR方法和RT-PCR进行了验证,从而获得了nuc1基因缺失突变株RN4220Δnuc1。
Two ORFs in Staphylococcus aureus genomes were predicted to encode nucleases. One is encoded the nuclease A (SNase) without a gene name (termed nuc1) ; the other is encoded a thermonuclease (TNase) with a name of nuc (termed nuc2 ). To further investigate the function of nuc2 in S. aureus, the resulting plasmid, pBT2 △nucl was successfully constructed. The recombinant plasmid pBT2 △nucl was introduced into S. aureus RN4220 by electroporation. Using the resistant marker for seven passages of culture, the recombination rate was about 2% (7/345). A nucl-deleted mutant of S. aureus strain RN4220 (termed RN4220△nucl ) was successfully constructed and conformed by PCR and RT-PCR.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2008年第11期43-47,共5页
China Biotechnology
基金
“十一五”国家科技支撑计划“食品安全关键技术”重大项目(2006BAK02A14)
上海市重大专项(07dz19508)
上海市科委平台建设项目(06dz22208)资助项目