| 耐甲氧西林葡萄球菌PBP2a的质粒克隆与构建 |
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| 引用本文： | 聂红,王云英,陈维贤.耐甲氧西林葡萄球菌PBP2a的质粒克隆与构建[J].中国微生态学杂志,2011,23(5):407-409. |
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| 作者姓名： | 聂红 王云英 陈维贤 |
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| 作者单位： | 重庆医科大学附属第二医院检验科,重庆,400010 |
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| 基金项目： | 重庆市卫生局医学科学技术研究项目 |
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| 摘 要： | 目的克隆并构建耐甲氧西林金黄色葡萄球菌（MRSA）青霉素结合蛋白2a（PBP2a）全长及转肽酶区的原核表达质粒。方法登录基因文库查找获得mecA基因的编码序列,应用PCR技术扩增获得DNA片段,将此基因片段插入PET-32a载体,同时酶切鉴定阳性克隆,DNA序列测定验证序列正确性。结果 PCR扩增获得了mecA基因全长及转肽酶区DNA片段,成功插入到原核表达载体PET32a,双酶切鉴定及DNA序列测定证实插入片段正确。结论成功构建了PBP2a全长及转肽酶区片段表达质粒,为该蛋白的纯化表达和疫苗研究奠定了基础。
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| 关 键 词： | 耐甲氧西林金黄色葡萄球菌 青霉素结合蛋白2a 转肽酶区 mecA基因 |
Clone and construction of prokaryotic expression plasmid of MRSA-PBP2a |
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| NIE Hong,WANG Yun-ying,CHEN Wei-xian.Clone and construction of prokaryotic expression plasmid of MRSA-PBP2a[J].Chinese Journal of Microecology,2011,23(5):407-409. |
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| Authors: | NIE Hong WANG Yun-ying CHEN Wei-xian |
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| Institution: | NIE Hong,WANG Yun-ying,CHEN Wei-xian(Department of Clinical Laboratory,the Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China) |
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| Abstract: | Objective To clone and construct the prokaryotic expression plasmid of penicillin binding protein 2a(PBP2a) of methicillin resistant Staphylococcus aureus(MRSA).Method According to the sequence of mecA gene published in GenBank,the target gene fragments were amplified by PCR,and then inserted into PET-32a plasmid.The recombined plasmids were identified by enzyme digestion and the inserted fragments were further confirmed by DNA sequencing.Result The corresponding prokaryotic expression plasmids of PBP2a had... |
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| Keywords: | Methicillin-resistant Staphylococcus aureus Penicillin binding protein2a Transpeptidase domain mecA gene |
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