三大不同品种马mtDNA Cytb基因PCR-RFLP分析

PCR-RFLP Analysis of the mtDNA Cytb Gene in Three Different Horse Breeds

用BamHⅠ、TaqⅠ、HaeⅢ、RsaⅠ和HincⅡ5种限制性内切酶通过PCR-RFLP技术检测了包括引入品种、培育品种和地方品种的6个类型(纯血马、三河马、乌珠穆沁马、锡尼河马、乌审马和矮马)共256匹马的mtDNACytb基因部分序列多态性。用8%非变性聚丙烯酰胺凝胶电泳将酶切产物分离,并用银染法显色。结果BamHⅠ和TaqⅠ表现出多态,5种酶共检测到7种态型,归纳为3种单倍型,以单倍型Ⅰ和Ⅲ为主体单倍型,但通过一个特殊的酶型BamHⅠ-B分析推测所研究的马可能起源于一个母系祖先。

Restriction endonucleases, namely BamH Ⅰ , Taq Ⅰ , Hae Ⅲ, Rsa Ⅰ and Hinc Ⅱ, were used to analyze the polymorphism of partial mtDNA Cytb gene sequences from 256 horses 6 types（Thoroughbred, Sanhe, Wuzhumuqin, Xinihe, Wushen and Pony） including the imported breed, cultivated breed and local breed. The products of endonuclease digestion were run on 8% non-denaturing polyacrylamide gel electrophoresis and detected by silver staining. Results indicated BamH Ⅰ and Taq Ⅰ polymorphism. In all,7 restriction patterns were defected that could be sorted into 3 haplotypes, of which haplotypes Ⅰ and Ⅲ were the basic haplotypes. We infer that these horses came from one female ancestor possibly through the analysis thorough one pattern, namely BamH Ⅰ -B.