CAJ | 학술논문

[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1×107,插入基因片断的大小为750～3000bp.,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.
[목적]구건미주구충cDNA고종이획득해구충적기인결구신식이편종중심조목적기인.[방법]종미주구충제삼기유충중제취mRNA,이mRNA위모판합성cDNA병련접입재체λZAPⅡ이구건cDNA고,수궤종해cDNA고중도선단극륭진행측서획득EST병여기인고중현유기인진행비교추도기편마단백.[결과]성공지구건료미주구충cDNA고,미확증고적적도위1×107,삽입기인편단적대소위750～3000bp.,고적중조솔고,병종고중획득11개EST서렬,기중7개여현유기인고중적모사공능단백유동원성.[결론]수차구건료대표성교고적미주구충cDNA고병종중감정출모사공능기인.
[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.Expresed se-quence tages (ESTs) were obtained by single pass sequencing of randomly isolated Cdna clones from the es-tablished library.[Results] A Cdna librazy of N.americanus was successfully constructed with high recombi-nant efficiency.The titer of unamplified library was 1×107.The insert size was about 750～3000bp.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 ome functional genes were identified from the library by ESTs.