喹乙醇致线粒体DNA基因损伤的SSCP分析  

Analysis of Polymerase Chain Reaction-Single Stranded Conformation Polymorphism on Olaquindox-Induced mtDNA Gene Damage in Chicken

作　　者：王德成[1] 高洪[1] 范泉水[2] 高斌[3] 

机构地区：[1]云南农业大学动物科技学院,昆明650201 [2]成都军区后勤部军事医学研究所,昆明650032 [3]云南农业大学食品科技学院,昆明650201

出　　处：《吉林农业大学学报》2007年第1期95-101,共7页Journal of Jilin Agricultural University

基　　金：云南省自然科学基金资助项目(2003C0049M)

摘　　要：为研究喹乙醇对mtDNA基因的致突变效应,以1日龄的健康艾维因商品代肉鸡为试材,运用改进的碱变性法提取肝脏的mtDNA,设计4对引物扩增mtDNA的tRNALeu(UUR)t、RNAIlet、RNALys、ATPase8 4个基因,同时运用SSCP技术对扩增产物进行突变分析。结果表明:喹乙醇处理Ⅵ组tRNALeu(UUR)有突变发生,序列测定结果表明该突变为4 007位的C→A点突变,其他各组的tRNALeu(UUR)基因无任何突变发生。该结果提示喹乙醇中毒可以导致mtDNA tRNALeu(UUR)基因突变。The experiment was made for investigating the mutational effect of Olanquindox-induced mtDNA gene, with 1-day healthy avian chicken as the materials. The liver mtDNA was extracted with an improved isolation of alkaline lysis, and detecting the mutations of tRNA^Leu（UUR）, tRNA^Ile, tRNA^Lys, ATPase8 genes by polymerase chain reaction-single stranded conformation polymorphism （PCR-SSCP） technique. It was found that a point mutation happened on the tRNA^Leu（UUR） gene in olaquindox-treated group- Ⅳ. The sequencing results showed this point mutation occurred in the position of 4 007, it was a point mutation of C→A transition. Simultaneously , no other mutations on this gene were detected in olaquindox-treated group. These results indicated that Olaquindox can cause the mutation on mtDNA tRNA^Leu （UUR） gene.

关 键 词：喹乙醇 聚合酶链式反应-单链构象多态性 线粒体DNA 基因损伤 

分 类 号：Q244[生物学—细胞生物学] Q523