应用长PCR扩增蝗虫线粒体全基因组

Amplification of Grasshoppers Complete Mitochondrial Genomes Using Long PCR

介绍了用两对长PCR引物扩增蝗虫(Acridoidea)线粒体全基因组的方法。从NCBI的核酸数据库下载得到36种已测昆虫线粒体全基因组,选取cytochromeb(Cytb)c、ytochrome oxidase subunitⅡ(COⅡ)、cytochrome oxidase subunitⅠ(COⅠ)基因的保守区域设计两对引物。其中引物LP03和LP04从COⅠ向Cytb扩增;引物LPCytb和LPCOⅡ从Cytb向COⅡ扩增,两对引物扩增的片段之间有大约1 kb的重叠。应用这两对引物成功扩增出10种蝗虫的线粒体基因组。考虑到在设计引物过程中所选序列在其他昆虫中的保守性,它们应能在大部分昆虫线粒体基因组扩增中发挥作用。

Long PCR was used to amplify complete mitochondrial genomes of 10 Acridoidea species. Two sets of primers were designed based on conserved regions of Cyt b, CO Ⅱ , and CO Ⅰ genes which were downloaded from the NCBI.One pair of primers（LP03 and LP04） amplified the region from CO Ⅰ to Cyt b ,while the other （LPCyt b and LPCO Ⅱ ） amplified the region from Cyt b to CO Ⅱ There was about 1 kb overlapping region between the two fragments. Although the primers used in this study were examined only in grasshoppers, a possibility of their application to other insects is very high, since the high degree of sequence conservation is shown on primer sites in other insects.