太平洋牡蛎核糖体DNA转录间隔子和线粒体基因片段序列测定

以相应引物经PCR扩增了太平洋牡蛎 (Crassostreagigas)的核糖体转录间区域 (ITS 1和ITS 2 )及线粒体 16SrDNA和COI基因片段。PCR产物经T 载体连接后进行克隆和测序 ,分别得到长度为 5 4 3、791、5 30和 70 0bp的核苷酸序列。 4个DNA片段的A、T、G和C碱基含量分别为 2 3.5 7%、2 0 .0 7%、2 9.4 7%和 2 6 .89% (ITS 1) ,2 7.4 3%、19.2 2 %、2 7.0 5 %和2 6 .30 % (ITS 2 ) ,2 9.2 5 %、2 9.2 5 %、2 3.0 2 %和 18.4 9% (16SrDNA) ,2 2 .71%、39.4 3%、2 0 .4 3%和 17.4 3% (COI)。实验证明ITS 1和ITS 2引物在贝类中通用性良好。文中同时讨论了 4个序列在我国几种牡蛎的种类鉴别及相关研究的应用潜力

Sequencing of ribosomal internal transcribed spacer regions and mitochondrial gene fragments in Crassostrea gigas

The ribosomal internal transcribed spacer regions (ITS 1 and ITS 2) and mitochondrial gene fragments ( 16S rDNA and COI) of Crassostrea gigas were amplified via PCR, and the PCR products were ligated into T vectors, cloned and sequenced.The nucleotide sequences of 543,791,530 and 700 bp from ITS 1,ITS 2,partial 16S rRNA gene and partial COI gene were obtained, respectively. The contents of A, T, G and C were 23.57%, 20.07%, 29.47% and 26.89% in ITS 1, 27.43%, 19.22%, 27.05% and 26.30% in ITS 2, 29.25%, 29.25% 23.02% and 18.49% in 16S rDNA, and 22.71%, 39.43%, 20.43% and 17.43% in COI in turn. The primers of ITS 1 and ITS 2 proved to be very universal in a variety of mollusk species. The potential uses of these four sequences for species identification and relevant research of several closely related oyster species were discussed.