中国动物传染病学报Issue(6)：36-42,7.
柔嫩艾美耳球虫子孢子酵母双杂交cDNA文库的构建
CONSTRUCTION OF A YEAST TWO-HYBRID CDNA LIBRARY FROM SPOROZOITES OF EIMERIATENELLA
摘要
Abstract
In order to screen the proteins related to the invasion of Eimeria tenella sporozoites, the yeast two-hybrid cDNA library of E.tenella sporozoites was constructed in the present study. The total RNA was isolated from the sporozoites and used as the template to synthesize the ifrst-strand cDNAs. The dscDNAs were acquired by long-distance polymerase chain reaction (LD-PCR) using the ifrst-strand cDNAs as the template and purified with Chroma Spin TE-400 Column to remove less than 200 bp fragments. The dscDNA,pGADT7-Rec and CarrierDNA were co-transformed into Y187 yeast competent cells. The dscDNA were then connected with pGADT7-Rec by homologous recombination reaction to construct the yeast two-hybrid cDNA library of E. tenella sporozoites. The results showed that the conversion ratio and titer of the cDNA library were 4.33×105/μg pGADT7-Rec and 3.62×107cfu/mL, respectively. As demonstrated in PCR ampliifcation, the cDNA library contained approximately 93.75%recombinant clones and the inserted cDNA fragments were between 200 bp and 2000 bp. Furthermore, 5 gene fragments were obtained by PCR amplication using the cDNA library as template and 5 speciifc pairs of primers of E.tenella. Therefore, it concluded that a yeast two-hybrid cDNA library of the sporozoites was constructed for screening invasion-related interaction proteins of the sporozoites of E.tenella.关键词
柔嫩艾美耳球虫/子孢子/酵母双杂交cDNA文库/同源重组Key words
Eimeriatenella/sporozoite/yeast two-hybrid cDNA library/homologous recombination分类
农业科技引用本文复制引用
徐帅兵,韩红玉,黄兵,赵其平,董辉,朱顺海,崔晓霞,谢雨翔,唐敏,杨志远..柔嫩艾美耳球虫子孢子酵母双杂交cDNA文库的构建[J].中国动物传染病学报,2016,(6):36-42,7.基金项目
国家自然科学基金（31272557，31572266）；农业部中央级公益性科研院所项目 （）
