中华哲水蚤线粒体DNA COI基因序列分析

Mitochodrial DNA COI Sequence Analysis of Calanus sinicus (Copepod) in Qingdao Waters

采用苯酚 氯仿 异戊醇(25∶24∶1)提取、异丙醇沉淀、酒精洗涤、TE缓冲液保存方法提取了采自胶州湾青岛海域的中华哲水蚤基因组DNA;以相应引物经PCR扩增得到线粒体DNA细胞色素氧化酶 I亚基基因(mtCOI)片段;PCR产物采用化学法进行质粒重组(载体pGEM T Easy Vector)、热休克转化重组质粒至大肠杆菌感受态细胞(JM109)、氨苄LB培养基扩大培养;测序.结果表明,中华哲水蚤线粒体COI碱基大小为710 bp(国际基因库索引号AY665663),其碱基组成A、C、G、T含量分别为24.23%、19.15%、22.54%和34.08%;G+C含量为41.69%,A+T为58.31%.

Calanus sinicus is a dominant species in the zooplankton communities of the Yellow Sea,Bohai Sea and the East China Sea, and is important food for many pelagic fishes especially their larvae.The specimens in the present study were collected from Qingdao Waters in May of 2002,by horizontal haul.The genomic DNA of Calanus sinicus was extracted by phenol:Chloroform:isoamyl alcohol (25241) and receipted by isopropyl alcohol.Mitochondrial DNA COI gene fragments were amplified successfully under our PCR conditions,then were recombined with pGEM-T Easy Vector (Promega Co.) and sequenced.The length of the sequence (accession no.AY665663 in GenBank) was 710 base pairs,and its composition (A,C,G and T) was 24.23%、19.15%、(22.54%)和34.08%, respectively.Alignment was completed between the mtCOI sequence collected from Qiaodao Waters and from Inland Sea,Japan (accession no.AF332769,GenBank),and the identity by calculated with DNAMAN software is 99.53%.The correctness and significance of nucleotide sequence met in our study was discussed in the present paper.