不同地理株旋毛虫细胞色素氧化酶Ⅰ基因多态性的PCR-单链构象多态性分析

PCR-SSCP analysis on partial mitochondrial cytochrome c-oxidase subunit Ⅰ (COXⅠ) genes of different Trichinella spiralis isolates

目的:观察不同地理株旋毛虫细胞色素氧化酶Ⅰ(COXⅠ)基因片段的多态性。方法:根据旋毛虫mtD-NACOXⅠ设计引物,应用PCR-单链构象多态性(PCR-SSCP)技术对我国7个猪源旋毛虫(Trichinella spiralis,T1)地理株(河南、湖北、云南、西安、天津、黑龙江同江及哈尔滨株)与1个波兰猪源旋毛虫(T1,编号ISS3)地理株进行COXⅠ基因片段多态性进行分析。结果:我国7个猪源旋毛虫地理株与波兰地理株COXⅠ基因均扩增出约400bp的片段,PCR扩增产物经SSCP检测发现有3种基因型(AA、AB和BB),波兰地理株为AA型,黑龙江同江、哈尔滨、西安、云南、湖北及河南株均为AB型,天津株为BB型,其中以AB基因型频率最高(0.75),AA与BB基因型均为0.125;A和B等位基因频率均为0.5;旋毛虫不同地理株COXⅠ基因的多态性信息含量为0.375,为中度多态性。结论:8个不同地理株猪源旋毛虫的COXⅠ基因为中度多态性。

Aim： To observe the polymorphism of the mitochondrial cytochrome c-oxidase subunit Igene of different I. spralis isolates. Methods：PCR-single-strand conformation polymorphism （PCR-SSCP） was used to analyze the polymorphism of the partial mitochondrial cytochrome c-oxidase subunit I gene （COX I ） of seven swine＇T, spiralis（T1 ） isolates from Henan, Yunnan, Harbin,Tougjiang in Heilongjiang, Hubei and Tianjin as well as one Poland isolate from domestic pig. Results： The results of electrophoresis of PCR products of Trichinella COX I gene showed that the band （400 bp） of seven Trichinella isolates from China was the same as Poland isolate. Three genotypes （AA, AB and BB） among different TrichineUa isolates were observed by SSCP. Poland isolate was AA genotype, Tianjin was BB geuotype, other isolates were AB geootype. The frequency of AB genotype was the highest （0.75） , the frequency of AA and BB genotype was 0. 125, and the frequency of A allele was equal to B allele （0.5）. The polymorphism information content （PIC） of different Trichinella isolates was 0. 375. Conclusion： The polymorphism of the COX I gene of eight swine Trichinella isolates was moderate.