建立猪线粒体ＤＮＡ Ｄ－ｌｏｏｐ５‘端和３’端ＰＣＲ扩增方法。方法：设计三对引物，应用ＰＣＲ对西双版纳近交系小耳猪，广西巴马小型猪，贵州小型香猪，大约克夏猪，荣昌猪和长白猪血液总ＤＮＡ样品中ｍｔＮＤＡ Ｄ－ｌｏｏｐ，５‘端进行扩增，琼脂糖凝胶电泳检测，结果：三对引物扩增带清晰，与已知片段长度一致。结论：引物设计合理，扩增方法可行，为进一步应用ｍｔＤＮＡ Ｄ－ｌｏｏｐ ＰＣＲ－ＰＲＬＰ，５’端ＳＳＰ
Objective: To establish the PCR technique for the amplification of the D loop and its 5' and 3' ends of porcine mitochondrial DNA (mtDNA). Methods: Three pairs of primers were designed to amplify the D loop and its 5' and 3' ends of mtDNA of Xishuangbanna small ear pig of the inbred stain, Guizhou Miniature fragrant pig, Guangxi Bama miniature pig, Rongchang pig, large Yorkshire pig and Landrace pig. The amplification products were detected with electrophoresis on 1.2% agarose gel. Results: Clearly visible amplification bands were obtained and the fragment length was the same as that reported previously. Conclusion: Our findings suggest that the design of the primers is reasonable and the procedures are reliable. The PCR technique we have established can be used in PCR RFLP to study of D loop of mtDNA, in SSCP to study the 5' and 3' ends of the D loop of mtDNA, in SSCP to study the 5' and 3' ends of the D loop and in direct sequencing.
Acta Academiae Medicinae Militaris Tertiae