大黄鱼16S rRNA和18S rRNA基因的测定与序列分析  

Cloning and Sequence Analysis of 16S rRNA and 18S rRNA Genes Fragments of Larimichthys crocea

作　　者：卢韫[1] 张际峰[1] 卢诗瑶 陈文明[1] 

机构地区：[1]淮南师范学院生命科学系,安徽淮南232001 [2]安徽科技工程学院生物化学工程系,安徽芜湖241000

出　　处：《激光生物学报》2010年第4期458-465,共8页Acta Laser Biology Sinica

基　　金：安徽省教育厅自然科学研究项目(KJ2010B443;KJ2009B268Z);淮南师范学院校级质量工程建设项目(JPKC200803)

摘　　要：利用多对引物,扩增并测定出大黄鱼16S rRNA基因和18S rRNA基因的部分序列,其长度分别为1 202 bp和1 275 bp,16S rRNA基因序列的GC含量为46.12%,18S rRNA基因的GC含量为53.00%。将大黄鱼16S rRNA基因序列与GenBank中15种硬骨鱼类的同源序列结合,同时将其18S rRNA基因序列与GenBank中9种脊索动物的同源序列相结合,运用软件获得各自序列间差异百分比,转换和颠换数值等信息。基于这两种基因序列,利用NJ法和BI法,分别构建16种硬骨鱼类和10种脊索动物的分子系统树。18S rRNA构建的系统树包括三大支,一支为哺乳类、鸟类和爬行类共6个物种,一支为两栖类的1个物种,另一支为2种硬骨鱼类。16S rRNA构建的系统树显示大黄鱼所在的石首鱼科与鲈科和盖刺鱼科亲缘关系较近。此外还讨论了这两个基因的序列特征。The partial nucleotide sequences of 16S rRNA gene and 18S rRNA gene were amplified and determined for Larimichthys crocea by PCR with several pairs of specific primers. The nt sequences of 16S and 18S rRNA genes of L. crocea were 1 202 bp and 1 275 bp in length, respectively. The G ＋ C content of 16S rRNA gene was 46.12 % , and the G ＋ C content of 18S rRNA gene was 53.00 %. The newly determined sequence of 16S rRNA gene ofL. crocea was analyzed together with homologous sequences of 15 fish species available in the GenBank database, and the determined sequence for 18S rRNA gene was combined with homologous sequences of 9 Chordate Species downloaded from Gen- Bank. The percentage nucleotide sequence divergence, and base substitutions were analyzed with the software. Based on 18S rRNA genes sequences, molecular phylogenetic trees constructed by neighbor-joining（ N J） and Bayesian （BI） methods supported the same topology, in which 9 species of vertebrates could be divided into three major monophyletic groups. The first group contained 6 species of amniotes. The second group comprised 1 species, Bufo guttatus, the member of Amphibians （ class Amphibia）. The remaining 2 species of fish belonged to the third group, with a clade comprising L. crocea, and Scophthalmus maximus. Another phylogenetic tree for 16 sequences of fish were derived using 16S rRNA gene and the same analytical approaches： NJ and BI, and Diplophos taenia（ AB034825 ） and Sigmops gracilis （AB016274） were used the outgroup species. The present results indicated that Sciaenidae was more closely related to Pomacanthidae and Percidae within Perciformes. In addition, several characteristics of 16S and 18S rRNA genes sequences were discussed in vertebrates.

关 键 词：大黄鱼 16S RRNA基因 18S RRNA基因 序列分析 

分 类 号：Q78[生物学—分子生物学]