中文摘要：

为研究吕氏泰勒虫入侵绵羊淋巴细胞的机理,用绵羊外周血淋巴细胞构建酵母双杂交文库,筛选与TlSP蛋白互作的蛋白。以密度梯度离心法分离出未感染泰勒虫的绵羊淋巴细胞,TRIzol法提取总RNA,建立酵母双杂交cDNA文库。结果显示,对文库进行评定,细胞密度为1.075×109/mL,插入片段长度在250~3 000bp之间,文库滴度为1.088×108 CFU,重组率为92%。利用诱饵质粒pGBKT7-TlSP对文库进行筛选,结果筛选到9种蛋白。生物信息学分析结果显示,这些蛋白主要参与调控基因转录、mRNA翻译、细胞的增殖、分化和凋亡等过程。上述结果表明,通过酵母双杂交技术筛选出与TlSP互作的蛋白,据此推测TlSP蛋白可能与吕氏泰勒虫在绵羊淋巴细胞内的增殖相关。

英文摘要：

The function of TlSP protein about invasion was studied through screening of the TlSP interacted proteins by the yeast two-hybrid system.The lymphocytes were isolated from uninfected sheep with protozoan by density gradient centrifuge,the total RNA was extracted from these cells by TRIzol reagent,and an yeast hybrid cDNA library was established.In result,the library evaluation shows that the cell density was 1.075×109 cell/mL,the insert fragments were between 250 bp and 3 000 bp and an average size of insert of the capacity of the library was 1.088×108 CFU with a recombinant rate of 92%.Nine proteins were obtained by using yeast two-hybrid system,and the bioinformatics analysis showed that these proteins were involved in regulating gene transcription,mRNA translation,cell proliferation,differentiation and apoptosis.In conclusion,the TISP interacted proteins screened by yeast two-hybrid may be involved in replication of T.luwenshuni in ovine lymphocyte.