柔嫩艾美耳球虫马杜拉霉素抗药株与敏感株的mRNA差异显示

利用柔嫩艾美耳球虫马杜霉素敏感虫株裂殖子和抗药虫株裂殖子为材料,对马杜霉素敏感虫株和抗药虫株进行差异显示PCR,共回收34条电泳差异片段,反向Northern点杂交鉴定4个片段为真正差异片段,并对4个片段进行测序、B1ast同源性比较.来自于马杜霉素抗药虫株裂殖子的ACD3-2序列与柔嫩艾美耳球虫微线-5同源性99%,说明AcD3-2序列是该基因的部分序列,微线-5蛋白与虫体融解宿主细胞膜、入侵、运动和溢出有关,在第二代裂殖子时期,抗药性虫株该序列发生转录,而在敏感虫株中沉默;HCD1序列来自马杜霉素敏感虫株,与柔嫩艾美耳球虫表面抗原16和17序列同源性分别为83%和86%,说明与这2个基因同源.AGD5片段来自抗药虫株,HAD8-2序列来自敏感虫株,通过比较这2条序列可能为新序列.

mRNA Differential Display PCR of Maduramisin Sensitive and Resistant Eimeria tenella Strain

The differential expression between merozoites of maduramisin sensitive and maduramisin resistant strains of E. tenella was detected with the technique of random mRNA Differential Display. The 34 differential DNA fragments were identified by reverse Northern dot hybridization. Four fragments were found to be really different, the differential expression fragments were cloned into pMD-18T vector and sequenced. Then the possible functions and the relationship of the 4 differential DNA fragments to drug resistance were predicted with Blast. Fragment ACD3-2 was found to be similar to Eimeia tenella microneme-5 gene, which is related to the parasite invasion, moving; Fragment HCD1 similar to part sequence of Eimeia tenella surface antigen-17, 16 genes, the other 2 differential expression sequences maybe new genes because no homologous gene were found in GenBank by Blast analysis.