摘要
为了建立可对鳕鱼DNA进行特异性检测的PCR方法,应用在出口贸易中的鳕鱼疑似样品和保护执法工作中难以检查辨认的鳕鱼产品的物种鉴定,从GenBank数据库下载7个鳕属及其他5种鱼类的mtDNA细胞氧化酶Ⅱ(COXⅡ)基因序列,并用DNA Star7.0软件对上述不同鳕鱼的该基因碱基序列进行比较。在此基础上,综合考虑了设计引物的基本原则,选择了鳕鱼与其他鱼类碱基序列上差异位点较多的两个片段,设计出PCR引物。用该引物分别对从7种鳕鱼和5种非鳕鱼的肌肉、脏器组织中提取的DNA进行PCR扩增。结果表明,所设计的引物对鳕鱼DNA具有特异性,从而达到了对该物种进行特异性检测鉴定的目的。
In order to develop a PCR method specifically amplifying DNA of ling species for identification of doubtful samples or unable recognized products in export commerce or forensic tests,mtDNA COX2 sequences of 7 ling species and 5 other fish species were download from GenBank,and compared by DNASTAR7.0 software.Considering the essential doctrine of designing PCR primers,two regions in which all ling share an unique hyplotype against other species were selected to design a pair of primers（Primer 1,2） to amplify a ling COX2 fragment specifically.PCR tests were performed with DNAs extracted from muscles,visceral tissues of 7 ling species and other fish species using this primer set.The results indicated that the primers were specific to ling DNA,and the identification of ling species by this method was sensitive,reproducible and reliable.
出处
《动物医学进展》
CSCD
北大核心
2011年第2期19-24,共6页
Progress In Veterinary Medicine
基金
山东检验检疫局科研目计划(SK200906)
