柔嫩艾美耳球虫第二代裂殖子中的Etmic-2基因克隆及测序  

Cloning and sequence of Etmic-2 gene form second generation of merizoites of Eimeria tenella

作　　者：蒋建林[1] 蒋金书[1] 

机构地区：[1]中国农业大学,北京海淀100094

出　　处：《中国兽医杂志》2002年第3期7-10,共4页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金资助项目 ( 30 170 6 95 )

摘　　要：作者根据子孢子微线基因 (Etmic- 2 )的 c DNA序列 ,利用计算机设计出 1对引物 ,从柔嫩艾美耳球虫第二代裂殖子中用 RT- PCR技术扩增出 1个片段 (m ic2 - m z)。把这个片段克隆到 p GEM- T- Easy Vector中后 ,我们得到 1个阳性克隆 pmic2 - mz,酶切分析证明 pm ic2 - m z含有 m ic2 - m z,并且外源片段以反方向插入 T载体中。核苷酸序列测定结果表明 m ic2 - m z与 Etmic- 2基因的编码区序列一样长 ,分子量都为 112 7bp,并且两序列之间也只有 3个碱基不同 ,也没有造成氨基酸编码错位 ,这说明球虫从子孢子到第二代裂殖子的二次无性生殖过程中 ,微线基因Etm ic- 2 c NDA序列没有发生太大的变化。According to the Etmic-2 cDNA sequence published by Tomley,a pair of primers were designed by using computer software.A 1.1kb cDNA encoding sequence of Etmic-2 from second generation of merizoites,named mic2-mz,was amplified by RT-PCR taking cDNA as template.This PCR product was cloned into pGEM-T-Easy vector by T-A ligation,and identified with restriction endonuclease analysis,and sequenced by automatic DNA sequencer.Comparing and analysis sequence of mic2-mz with that of Etmic-2indicates two gene have same molecular weight,and mic2-mz also only has three nucleotide change with Etmic-2,and this three changed nucleotide don't interrupt open reading frame.This implies that during parasite development cycle from oocysts to second generation merizoites Etmic-2 gene don't change.

关 键 词：Etmic-2基因 克隆 测序 柔嫩艾美耳球虫 微线基因 第二代裂殖子 

分 类 号：S852.723[农业科学—基础兽医学]