应用长PCR扩增蝗虫线粒体全基因组  被引量:19

Amplification of Grasshoppers Complete Mitochondrial Genomes Using Long PCR

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作  者:刘念[1] 胡婧[1] 黄原[1] 

机构地区:[1]陕西师范大学生命科学学院,西安710062

出  处:《动物学杂志》2006年第2期61-65,共5页Chinese Journal of Zoology

基  金:国家自然科学基金资助项目(No.30470238)

摘  要: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.

关 键 词:长PCR 引物 蝗虫 线粒体基因组 

分 类 号:Q78[生物学—分子生物学]

 

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