›› 2003, Vol. 23 ›› Issue (1): 3-6.

所属专题: 纪念实验动物学前辈-李厚达老师专辑

• 论文 • 上一篇    下一篇

金黄色葡萄球菌耐热核酸酶nuc基因的克隆和序列测定

  

  1. 扬州大学比较医学中心,扬州 225009
  • 收稿日期:2002-03-12 出版日期:2003-01-25 发布日期:2013-03-19
  • 基金资助:

    “十五”国家科技攻关计划“国家遗传工程小鼠资源库的建立”(项目编号:2001BA70113)

Cloning and Sequencing of the nuc Gene of Staphylococcus aureus

  1. Comparative Medicine Center,Yangzhou University,Yangzhou 225009, China
  • Received:2002-03-12 Online:2003-01-25 Published:2013-03-19

摘要: 根据已公布的金黄色葡萄球菌耐热核酸酶nuc基因的序列,设计并合成一对特异性的引物,以鼠金黄色葡萄球菌临床分离株SA.m0109的基因组DNA为模板,利用PCR技术对其耐热核酸酶nuc基因片段进行扩增,获得了预计大小的片段。将这一片段克隆到质粒载体pGEM-T中,对重组质粒pGEM-NUC进行限制性内切酶分析、PCR鉴定和克隆片段的序列测定,证实了克隆片段的可靠性。

关键词: 金黄色葡萄球菌, 聚合酶链反应, 分子克隆, 序列测定

Abstract: According to the sequence of the nuc gene, which encodes the thermostable nuclease (TNase) of Staphylococcus aureusS. aureus), a pair of oligonucleotide primers which are special to the sequence of nuc gene were designed and synthesized. Using the genome DNA of the clinically isolated strain of S. aureus SA. m0109 as template, the fragment flanking nuc gene was amplified by Polymerase Chain Reaction (PCR). The expected size fragment was obtained and was cloned into the plasmid pGEM-T. The recombinant plasmid of pGEM-NUC was identified by restriction enzyme analysis, PCR and sequencing, which proved completely its validity.

Key words: Staphylococcus aureus, Polymerase Chain Reaction, Molecular clone, Sequencing