| 卡介苗阿拉伯糖苷转移酶embC基因敲除株的构建和初步鉴定 |
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| 引用本文： | 李娅莎,鲍朗,李岩,张会东,赵明才.卡介苗阿拉伯糖苷转移酶embC基因敲除株的构建和初步鉴定[J].中国微生态学杂志,2009,21(5):440-442. |
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| 作者姓名： | 李娅莎 鲍朗 李岩 张会东 赵明才 |
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| 作者单位： | 1. 重庆医科大学附属儿童医院,儿科研究所,重庆,400014
2. 四川大学,华西医学中心感染免疫室,四川,成都,610041 |
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| 摘 要： | 目的采用基因敲除技术构建了卡介苗embC基因缺失株。方法从卡介苗基因组中扩增出embC基因，定向插入自杀质粒p2NIL中，切除embC基因中约1000bp片段使其失活，再定向插入标记片段，筛选鉴定阳性克隆，电穿孔转入卡介苗，筛选重组菌株。结果PCR和酶切鉴定证明构建成功用于基因打靶的置换型自杀质粒，并筛选成功获得重组卡介苗。结论获得了卡介苗embC基因敲除株，为进一步研究对卡介苗免疫活性的影响奠定了基础。
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| 关 键 词： | 卡介苗 embC基因 基因打靶 同源重组 |
Construction and identification of Mycobacterium BCG strain with the mutant embC gene |
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| LI Ya-sha,BAO Lang,LI Yan,ZHANG Hui-dong,ZHAO Ming-cai.Construction and identification of Mycobacterium BCG strain with the mutant embC gene[J].Chinese Journal of Microecology,2009,21(5):440-442. |
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| Authors: | LI Ya-sha BAO Lang LI Yan ZHANG Hui-dong ZHAO Ming-cai |
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| Institution: | LI Ya-sha ,BAO Lang ,LI Yan ,ZHANG Hui-dong ,ZHAO Ming-cai ( 1. Pediatric Research Institute, Children＇s Hospital, Chongqing University of Medical Sciences, Chongqing 400014, China; 2 Department of Infection and Immunity, West China Medical Center, Sichuan University, Chengdu 610041, China ) |
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| Abstract: | Objective To construct the mutant embC gene of Mycobacterium BCG strain using gene knockout. Method The complete embC gene was amplified from BCG genome and inserted into the suicide plasmid p2NIL, then an inner fragment of about 1000 bp was cut down form the mutant of embC gene and the mark caste was engaged to constract the gene-target vector. The recombinant vector was transformed into Mycobacterium BCG strain by electroporation method. Result PCR and restriction endonuclease map revealed the successful construction of the suicide vector and the mutant strain. Conclusion The recombinant BCG strain which lost embC gene function was constructed, laying a foundation for further research on rBCG immune function. |
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| Keywords: | BCG embC gene Gene knockout Homologous recombination |
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