| 鸡致病性大肠杆菌O_1、O_2和O_(78)的Ι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： | 王文成,张贵刚.鸡致病性大肠杆菌O_1、O_2和O_(78)的Ι型菌毛结构基因的克隆、序列测定及同源性分析[J].中国兽药杂志,2002(12). |
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| 作者姓名： | 王文成 张贵刚 |
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| 作者单位： | 辽宁省益康生物制品厂 辽宁辽阳111000
(王文成)，辽宁省益康生物制品厂 辽宁辽阳111000(张贵刚) |
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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作为模板,用TD-PCR技术从其中分别扩增出了0.55kb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载体中,转化至受体菌JM109中,用Amp/IPTG/X-gal琼脂平板蓝白菌落筛选法,得到含阳性重组子的菌株,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549bp,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,此3菌株的I型菌毛基因的同源性为89.9%~92%。
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| 关 键 词： | 鸡致病性大肠杆菌 I型菌毛 TA克隆 DNA测序 同源性 |
Cloning and Sequence Analysis of Type 1 Pilin Structure Gene of Chicken Pathogenic Escherichia coli Serotype O_1, O_2 and O_(78) |
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| WANG Wen-cheng,ZHANG Gui-gang.Cloning and Sequence Analysis of Type 1 Pilin Structure Gene of Chicken Pathogenic Escherichia coli Serotype O_1, O_2 and O_(78)[J].Chinese Journal of Veterinary Drug,2002(12). |
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| Authors: | WANG Wen-cheng ZHANG Gui-gang |
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| Abstract: | Type 1 pilin structure gene (piliA) from chicken pathogenic Escherichia coli O1,O2 and O78 wasgot by polymerase chain reaction (PCR). PCR products were isolated by 1.5% agarose gel electrophoresisand recovered by PCR preps DNA purification system. Then they were inserted in pGEM-T vector. Bytransformation of JM109 and screening with Amp/IPTG/X-gal agar plate, recombinant strains JM109(pTFimO1, pTFimO2 and pTFimO78)were got. These recombinant plasmids pTFimO1, pTFimO2 and pTFimO78were studied in detail by restriction endonuclease analysis. The results showed that the recombinant plasmidscarried type 1 pilin structure gene. Homologous were compared and analyzed with the help of DNA Starprograms. Similarities of piliA gene were 89.9%~92%. |
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| Keywords: | chicken pathogenic Escherichia coli type 1 pilin TA cloning DNA sequence analysis homology |
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