贝氏莫尼茨绦虫成节及孕节间7个差异基因EST的克隆及序列分析  

Analysis and Cloning of Seven Differential ESTs Between Mature and Gravid Proglottids of Monie^ja benedeni

作　　者：罗盘棋[1,2] 王新华[1] 薄新文[1] 

机构地区：[1]新疆兵团绵羊繁育生物技术重点实验室,新疆石河子832000 [2]石河子大学动物科技学院,新疆石河子832003

出　　处：《寄生虫与医学昆虫学报》2009年第3期129-133,共5页Acta Parasitologica et Medica Entomologica Sinica

摘　　要：本研究以新疆绵羊体内寄生的贝氏莫尼茨绦虫为研究材料，运用mRNA差异显示方法，研究了贝氏莫尼茨绦虫成节和孕节节片间差异基因信息。提取贝氏莫尼茨绦虫成虫成节和孕节的总RNA为模板，以3条Oligo-dT（Ⅰ，Ⅱ，Ⅲ）锚定引物和8条随机引物配对的24种组合，进行DDRT-PCR扩增，经过8％聚丙烯酰胺凝胶电泳分离后以及银染方法，显示差异DNA条带。凝胶回收差异条带并进行再扩增，琼脂糖电泳检测，得到7个差异片段，经克隆测序及相似性比对，其中有6条均未见有报道，所以其应为新基因。Seq16与猪肉绦虫幼虫UNAM—cd2mRNA同源性达100％，UNAM-cd2 mRNA功能未知，与细粒棘球绦虫胃蛋白酶同源性达100％，与日本血吸虫G-10蛋白同源性达88％，G-10蛋白功能未知，与黑腹果蝇胚胎生殖腺同源性84％。Cestodes were stained with hematoxylin and observed under microscope. The total RNA as templet was extracted from both mature and gravid proglottids using mRNA differential display method. 24 combinations which matched 30ligo-dT （ Ⅰ ,Ⅱ , Ⅲ ） anchor primers and 8 random primers were used for DDRT-PCR amplification. The products were separated by 8 % polyacrylamide gel electrophoresis （PAGE） and stained with silver staining. The different bands were retrieved from agarose gel, amplified by PCR and detected by agarose electrophoresis, and a total of 7 different bands were obtained. The retrieved products were respec- tively cloned to pMD18-T and sequenced. These sequences were compared with that in international public bio-information databases. and then 6 new genes were confirmed. The homology of Seq16 with Taenia solium UNAM-cd2 mRNA and, Echinococcus gran- ulosus pepsinum, was 100% , with G-10 albumen of Schistosomajaponicum was 88%, and with embryo genital gland of Drosophila melanogaster was 84% .

关 键 词：DD—PCR 贝氏奠尼茨绦虫 基因克隆 序列分析 EST 

分 类 号：Q78[生物学—分子生物学] S858.27[农业科学—临床兽医学]