恶性疟原虫基因组文库的构建及未知抗原cDNA片段的筛选  

Construction of Plasmodium falciparum FCC1/HN genomic library and screening for a unknown antigenic cDNA fragment

作　　者：闫宗合[1] 谢毅[2] 毕惠祥[1] 李明[1] 吴小舟[2] 杨静 赵雨田 李英杰[1] 

机构地区：[1]第一军医大学热带医学研究所热带病研究室 [2]复旦大学遗传工程国家重点实验室,上海市200433

出　　处：《第一军医大学学报》1997年第2期94-97,共4页Journal of First Military Medical University

摘　　要：恶性疟原虫FCC1／HN株基因组DNA经Sau3AⅠ部分酶切，取14－23kb片段连接入GEW-11λ噬菌体置换型载体构建基因组文库。所得重组体数目为7.3×104pfu，近达构建完整恶性疟原虫基因组文库理论值的10倍。随机挑取该库10个噬菌斑，抽提λDNA，用XhoⅠ酶切，得到14－23kb的插入片段和载体双臂，符合建库要求．以抗体筛选恶性疟原虫cDNA表达文库所获得的未知抗原cDNA片段为探针，在其中筛选到了阳性克隆，为阐明该未知cDNA对应的基因奠定了基础．To construct a P. falciparum genomic library for genomic analysis of several unknown antigenic cDNA fragments aquired by screening of its cDNA expression library, the genomic DNA of FCC1/HN strain was partially cleaved with Sau3A Ⅰ,then the 14 to 23 kb fragments were fractionated and ligated into GEM-11 Lambda replacement vector. After packaged in vitro, 7. 3 ×104 recombinants were obtained,which were approximately 10 times than the calculated sufficient P. falciparum genomic library size. The Lambda DNA of ten random clones in the library was cleaved with Xho Ⅰ,which recognized the Xho Ⅰsite on the GEM-11 multiple cloning sites. And all cleaved fragments, including the inserted 14 to 23 kb fragments and the vector arms,conformed to our inference. Furthermore,using a 6OObp unknown antigenic cDNA fragment as probe,We screened the library by hybridization and got a positive clone. Mapping and sequencing of the coressponding region of the cDNA in the genomic clone are under way.

关 键 词：恶性疟 疟原虫 基因组文库 CDNA 片段 

分 类 号：R531.3[医药卫生—内科学] R382.31[医药卫生—临床医学]