| 鸡致病性大肠杆菌O_1、O_2和O_(78)Ⅰ型菌毛结构基因的克隆、序列测定及同源性分析 |
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| 引用本文： | 王忠山. 鸡致病性大肠杆菌O_1、O_2和O_(78)Ⅰ型菌毛结构基因的克隆、序列测定及同源性分析[J]. 福建畜牧兽医, 2009, 31(5): 10-13 |
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| 作者姓名： | 王忠山 |
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| 作者单位： | 福州大北农生物技术有限公司,福州,350014 |
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| 摘 要： | 提取鸡致病性大肠杆菌分离株O1、O2和O78的基因组DNA作模板,用TD-PCR技术从其中分别扩增出0.55 KB的I型菌毛结构基因(piliA).将扩增得到的piliA基因片段,用TA克隆的方法分别克隆进pGEM-T栽体中,转化至受体菌JM109中,用Amp/IPTG/X-gal琼脂平板蓝白菌落筛选法,得到舍阳性重组子的菌株,提取质粒用PstI单酶切及NcoI和PstI双酶切鉴定,结果证实,所构建的克隆质粒中均含有相应piliA基因.经DNA序列分析,其结构基因阅读框架大小为549 bp,但其中O1菌株Ⅰ型菌毛基因在第72位发生突变,有6个碱基插入.经DNAStar核酸分析软件分析,3个基因同源性为89.9%～92.0%.
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| 关 键 词： | 鸡致病性大肠杆菌 I型菌毛 TA克隆 同源性 |
Cloning and sequence analysis of type 1 pili structure gene of chicken pathogenic Escherichia coli serotype O_1,O_2 and O_(78) |
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| Wang Zhongshan. Cloning and sequence analysis of type 1 pili structure gene of chicken pathogenic Escherichia coli serotype O_1,O_2 and O_(78)[J]. Fujian Journal of Animal Husbandry and Veterinary medicine, 2009, 31(5): 10-13 |
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| Authors: | Wang Zhongshan |
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| Affiliation: | Wang Zhongshan(Fuzhou dabeinong biotechnology limited company,Fuzhou 350014) |
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| Abstract: | Type 1 pili structre gene(piliA) from chicken pathogenic Escherichia coli O1,O2 and O78 by Polymerase Chain Reaction(PCR),PCR products were isolated by 1% agarose gel electrophoresis and recovered by PCR Preps DNA Purification System,then They were inserted pGEM-T vector.By transformation of JM109 and screening with Amp/IPTG/X-gal agar plate,we got recombinant strains JM109(pTFimO1,pTFimO2 and pTFimO78).These recombinant plasmids pTFimO1,pTFimO2 and pTFimO78 were studied in detail by restriction endonucleas... |
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| Keywords: | chicken pathogenic Escherichia coli type 1 pili TA cloning homology |
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