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| CPA-LTB融合基因的构建与表达 | |
| 引用本文： | 李自青,许崇波,赵志军,许崇利,曾瑾.CPA-LTB融合基因的构建与表达[J].中国兽医学报,2007,27(4):507-510. |
| 作者姓名： | 李自青 许崇波 赵志军 许崇利 曾瑾 |
| 作者单位： | 1. 山西大同大学医学院,山西,大同,037008 2. 大连大学生物工程学院,辽宁,大连116622 3. 宁夏大学生命科学学院,宁夏,银川,750021 |
| 基金项目： | 教育部科学技术研究重点项目 |
| 摘 要： | 利用PCR技术，从A型产气英膜梭菌染色体和pEWD299中分别扩增出a毒素（CPA）基因和LTB基因，通过分离、纯化、内切酶酶切、连接和转化，构建了含CPA—LTB融合基因表达质粒的重组菌株BL21（DE3）（pCPA—LTB）。经酶切鉴定和核苷酸序列测定证实，构建的重组质粒pET—CPA含有CPA—LTB融合基因，其基因序列和阅读框架均正确。经EISA检测和SDS—PAGE分析，重组菌株表达的CPA—LTB融合蛋白能够被α、LT毒素抗体所识别。重组菌株BL21（DE3）（pCPA—LTB）经IPTG诱导后，融合蛋白CPA—LTB的表达量占菌体总蛋白相对含量的12.5％。 |
| 关 键 词： | 基因融合 基因表达 |
| 文章编号： | 1005-4545（2007）04-0507-04 |
| 修稿时间： | 2005-09-08 |
Construction and expression of CPA-LTB fusion gene |
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| LI Zi-qing,XU Chong-bo,ZHAO Zhi-jun,XU Chong-li,ZEN Jin.Construction and expression of CPA-LTB fusion gene[J].Chinese Journal of Veterinary Science,2007,27(4):507-510. | |
| Authors: | LI Zi-qing XU Chong-bo ZHAO Zhi-jun XU Chong-li ZEN Jin |
| Institution: | 1. College of Medicine, Datong University, Datong , Shanxi 037008, China ; 2. College of Bioengineering, Dalian University, Dalian ,Liaoning 116622 ,China ; 3. Life Science School ,Ningxia University , Yinchuan 750021 ,China |
| Abstract: | Alpha-toxin gene from chromosomal DNA of Clostridium perfringens type A and heat-labile toxin B unit gene from plasmid pEWD299 containing heat-labile toxin of Escherichia coli were amplified by PCR.PCR products were cleaved with restriction endonucleases and recovered.The recombinant plasmid pCPA-LTB containing CPA-LTB fusion gene was constructed by recombinant technique and then transformed into Escherichia coli BL21(DE3).The recombinant expression plasmid pCPA-LTB was studied in detail by restriction endonucleases analysis and nucleotide sequencing.The results showed that the recombinant expression pCPA-LTB possessed the positive alpha-toxin and heat-labile toxin B unit gene sequences and their reading frames.The CPA-LTB fusion proteins were expressed in recombinant strain BL21(DE3)(pCPA-LTB) with the expression level of the CPA-LTB fusion proteins to be about 12.5% of the total cellular protein by SDS-PAGE and thin-layer gel scanning analysis. |
| Keywords: | CPA-LTB |
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