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当前位置: 首页 > 文章 > 鸡柔嫩艾美耳球虫石家庄多重耐药虫株差异基因的表达 中国兽医学报 2013,33 (9) 1373-1377
Position: Home > Articles > Differential display gene expression of chicken Eimeria tenella multiple resistance strain isolated from Shijiazhuang Chinese Journal of Veterinary Science 2013,33 (9) 1373-1377

鸡柔嫩艾美耳球虫石家庄多重耐药虫株差异基因的表达

作  者：
李佩国;张艳英;贾青辉;张香斋;李蕴玉;张文香
单  位：
河北科技师范学院河北省预防兽医重点实验室
关键词：
鸡;柔嫩艾美耳球虫;差异显示PCR;变性聚丙烯酰胺凝胶电泳;基因克隆
摘  要：
采用Trizol试剂法提取鸡柔嫩艾美耳球虫石家庄多重耐药虫株的总RNA,利用3种锚定引物和20种随机引物进行RT-PCR,产物经变性聚丙烯酰胺凝胶电泳后银染,共回收10条差异条带,进行2次PCR扩增,产物回收后与pMDTM18-T Vector连接转化,再进行斑点杂交试验、序列分析和同源性比较。结果表明,石家庄多重耐药性虫株的mRNA的S116序列与GenBank和Sanger上已发表的柔嫩艾美耳球虫第1段染色体上882bp长的序列同源性达99%,为未知蛋白。这为克隆全长cDNA和探索球虫耐药性产生的分子机理奠定了基础。
译  名：
Differential display gene expression of chicken Eimeria tenella multiple resistance strain isolated from Shijiazhuang
作  者：
LI Pei-guo;ZHANG Yan-ying;JIA Qing-hui;ZHANG Xiang-zhai;LI Yun-yu;ZHANG Wen-xiang;Hebei Normal University of Science and Technology,Key Laborationary Veterinary Medicine in Hebei Province;
关键词：
chicken;;E.tenella;;mRNA differential display PCR;;denaturing polyacrylamide gel;;gene cloning
摘  要：
Total RNA of E.tenelladrug-resistant strain from Shijiazhuang was extracted with Trizol.DDRT-PCR was established by using 3 anchored primers and 20 arbitrary primers.The products of PCR were analyzed on the denaturing polyacrylamide gels by silver-straining.Ten differential fragments were excised from the gels and reampilfied with the same sets of primers.Then the PCR products were purified and ligated with pMDTM18-T vector.The Dot-blot hybridization proved that the one clone from Shijiazhuang drug-resistant strain of E.tenellaappeared the strong signals.After sequence analysis compared with homogenize,the similarity was 99% among the sequence S116 from mRNA of Shijiazhuang drug-resistant strain with the sequence 882 bp length in the first chromosome of E.tenellain GenBank and Sanger,and it was an unknown protein.This study paved the way for cloning the full-length cDNAs and finding the molecular mechanism about the drug-resistance of E.tenella.

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