【作者】 王德成； 高洪； 范泉水；

【Author】 WANG De-cheng GAO Hong FAN Quan-shui (College of Animal Science and Technology, Yunnan Agricultural University, kunming 650201.The Military Medical Institute of Chengdu Military Area, kunming 650032)

【机构】 云南农业大学动物科技学院； 成都军区联勤部军事医学研究所；

【摘要】 为了研究喹乙醇对线粒体DNA基因的致突变效应,将72只一日龄的健康艾维因商品代肉鸡随机分为2组(36只／组):Ⅰ组(对照组)和Ⅱ组(喹乙醇处理组),两组都饲喂不含喹乙醇的饲料,并按标准鸡场的程序进行免疫和管理。处理组按照14．48mg／kg体重灌服喹乙醇,每周按体重变化改变给药量,同时每次改变给药量前剖杀试验动物。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)、tRNAIle、 tRNALys、ATPase8四个基因,运用SSCP(single stranded conformation polymorphism)技术对扩增产物进行突变分析。同时运用分光光度计法对血清丙二醛(malondialdehyde,MDA) 含量进行测定。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4007位的C→A点突变,其它各组的tRNALeu(UUR)基因无任何突变发生。MDA含量测定表明Ⅱ组从第二周开始明显高于Ⅰ组(P<0．05或P<0．01),说明机体内有异常水平的FRs 产生。该结果提示喹乙醇中毒可以导致mtDNA tRNALeu(UUR)基因突变。更多还原

【Abstract】 The experiment was amid at investigating the mutational effect of Olanquindox-induced mtDNA gene, 72 healthy avian chicken were divided randomly into 2 groups ( 36 chickens/group): normal group(ab.C) and olaquindox-treated group (ab:T), the programme of immunity and management processed accordingly to the standard hennery. Olaquindox was given from mouth with 14.48mg/kg in term of weight in olaquindox-treated group. The given dosage was changed in pace with weight weekly. Experimental animal was dissected before changed the dosage. The liver mtDNA was extracted with-an improved isolation of alkaline lysis, and detecting the mutations of tRNALeu(UUR)、tRNAIle、tRNALys、ATPase8 genes by polymerase chain reaction-single stranded conformation polymorphism (PCR-SSCP) technique. It was found that a point mutation happened on the tRNALeu(UUR) gene in olaquindox-treated group-vi, the sequencing results showed this point mutation occurred in the position of 4007, it is a point mutation of C→A transition. Simultaneously , no other mutations on this gene was detected in olaquindox-treated group. The MDA levels in group Ⅱ were remarkably higher than those in group Ⅰ from the 2 week (P<0.05 or P<0.01), and there was no significant difference in MDA level at the first week between two groups(P>0.05), we can safely conclude that there was abnormal quantity of FRs, These results indicated that Olaquindox can cause the mutation on mtDNA tRNALeu(UUR) gene.更多还原