摘要
[目的 ]构建美洲钩虫cDNA库从而获得该钩虫的基因结构信息以便从中寻找目的基因。 [方法 ]从美洲钩虫第三期幼虫中提取mRNA ,以mRNA为模板合成cDNA并连接入载体λZAPII以构建cDNA库 ,随机从该cDNA库中挑选单克隆进行测序获得EST并与基因库中现有基因进行比较推导其编码蛋白。 [结果 ]成功地构建了美洲钩虫cDNA库 ,未扩增库的滴度为 1× 10 7,插入基因片断的大小为 75 0～ 30 0 0bp .,库的重组率高 ,并从库中获得 11个EST序列 ,其中 7个与现有基因库中的某些功能蛋白有同源性。 [结论 ]首次构建了代表性较高的美洲钩虫cDNA库并从中鉴定出某些功能基因。
Objective] To obtain the genetic information on Necator americanus and to search for the purpose genes.[Methods] mRNA was isolated from the third stage larvae of Necator americanus maintained in hamsters. Double strand cDNA was synthesized and ligated to λZAPII vector to construct the cDNA library. Expressed sequence tages (ESTs) were obtained by single pass sequencing of randomly isolated cDNA clones from the established library. [Results] A cDNA library of N americanus was successfully constructed with high recombinant efficiency. The titer of unamplified library was 1×10 7. The insert size was about 750～3 000 bp. Of 11 ESTs obtained from the library, 7 have a significant homology with certain functional genes.[Conclunsion] A high quality and high representative cDNA library of N.americanus was constructed at the first time and some functional genes were identified from the library by ESTs.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
2000年第1期26-29,共4页
Chinese Journal of Parasitology and Parasitic Diseases
基金
美国卫生研究所热带病研究中心资助
