摘要
目的:建立能够快速、准确地进行塔里木马鹿的基因鉴定方法。方法:采用通用引物对L1091/H1478调出3种不同马鹿的线粒体12S rRNA片段进行比对分析,根据差异片段设计能够鉴别出塔里木马鹿的特异性引物对进行PCR鉴别,并寻找出一个能够识别差异片段的限制性内切酶进行酶切鉴别。结果:使用设计的特异性引物对CEYS/CEYA进行PCR,只有塔里木马鹿的基因组DNA可以获得扩增条带,可以很好的鉴别出塔里木马鹿;限制性内切酶PshBⅠ可以识别塔里木马鹿与其它马鹿的差异序列,只有塔里木马鹿的12S rRNA片段可以被切成2个小片段,可以鉴别是否为塔里木马鹿。结论:本实验建立的特异性PCR及PCR-RFLP可以作为塔里木马鹿的基因鉴定方法。
Objective： To develop a rapid and effective method of gene identification for Tarim Red Deer（C.elaphus xanthopygus）.Method： The mitochondrial DNA（mtDNA） 12SrRNA fragments of three different Red Deer（Cervus elaphus） was gained by general primers L1091/H1478.We aligned and analyzed the sequence to find the distinctive site,based on which we designed a pair of allele-specific primers for diagnostic PCR,and on which we found a presence of polymorphic sites for restriction endonuclease.Results： The allele-specific primers CEYS/CEYA can only amplify the DNA fragment of DNA templates from Tarim Red Deer in PCR,and the PshBⅠ restriction endonuclease can only digest the 12SrRNA fragment from Tarim Red Deer into two smaller pieces.So these two techniques provide effective approaches to identify Tarim Red Deer.Conclusion： The methods of Allele-Specific Diagnostic PCR and PCR-RFLP developed in this equipment can provide accurate identification for Tarim Red Deer.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2011年第3期570-573,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
