| 血清Ⅱ型鸭疫里默氏杆菌基因组文库的构建 |
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| 引用本文： | 林树乾,何元龙,赵增成,李峰.血清Ⅱ型鸭疫里默氏杆菌基因组文库的构建[J].中国兽医寄生虫病,2010,18(6):38-42. |
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| 作者姓名： | 林树乾 何元龙 赵增成 李峰 |
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| 作者单位： | 山东省农业科学院家禽研究所,济南250023 |
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| 基金项目： | 山东省科学技术发展计划，高校院所自主创新计划 |
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| 摘 要： | 为研究血清Ⅱ型鸭疫里默氏杆菌的抗原基因,筛选毒力较强的血清Ⅱ型鸭疫里默氏杆菌菌株,提取其基因组,分别用Sau3A I酶切法和shotgun法对基因组进行处理后,选取1.5～3 kb的DNA片段,与PUC118载体连接后电转化JM109大肠杆菌感受态细胞,构建血清Ⅱ型鸭疫里默氏杆菌基因组文库。两种方法得到的库容量分别为40 000和50 000左右,随机挑选阳性克隆鉴定表明有外源DNA片段的插入,说明建库成功。该文库的成功构建为Ⅱ型鸭疫里默氏杆菌新的抗原基因的筛选奠定了基础。
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| 关 键 词： | 鸭疫里默氏杆菌 基因文库 构建 |
CONSTRUCTION OF GENOMIC LIBRARY OF RIEMERELLA ANATIPESTIFER SEROTYPE II |
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| LIN Shu-qian,HE Yuan-long,ZHAO Zeng-cheng,LI Feng.CONSTRUCTION OF GENOMIC LIBRARY OF RIEMERELLA ANATIPESTIFER SEROTYPE II[J].Chinese Journal of Veterinary Parasitology,2010,18(6):38-42. |
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| Authors: | LIN Shu-qian HE Yuan-long ZHAO Zeng-cheng LI Feng |
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| Institution: | (Poultry Institute of Shandong Academy of Agricultural Sciences,Jinan 250023,China) |
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| Abstract: | The genomic library of a virulent Riemerella anatipestifer serotype Ⅱ was constructed to identify the protective antigens.The genomic DNA was digested with restriction endonuclease Sau3A Ⅰ and cut into pieces by shotgun method.These DNA fragments with the size from 1.5 to 3 kb were ligated into the plasmid vector PUC118.Then,the recombinant plasmid was transformed into E.coli JM109 by electroporation to construct the genomic library with the capability about 40000 and 50000.The insertion of exotic DNA fragments was confirmed by PCR that amplified positive clones from random selected samples. |
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| Keywords: | Riemerella anatipestifer genomic library construction |
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