太湖新银鱼m tDNA COII和Cytb基因的克隆与序列分析  

Cloning and Sequence Analysis of Mitochondrial COII and Cytb Genes of Neosalanx taihuensis

作　　者：张际峰[1] 卢韫[1] 杜晓光[1] 汪承润[1] 聂刘旺[2] 王洋[2] 

机构地区：[1]淮南师范学院化学生物系,安徽淮南232001 [2]安徽师范大学生命科学学院,安徽芜湖241000

出　　处：《激光生物学报》2008年第2期229-234,共6页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(30370211);淮南师范学院青年教师科研启动基金项目(2005);淮南师范学院青年教师自然科学基金项目(2006LKQ03);安徽省教育厅青年教师资助项目(2007jq1163)

摘　　要：设计了5对特异性引物,扩增、拼接并测定出太湖新银鱼线粒体tRNAAsp-COII-tRNALys和tRNAGlu-Cytb-tRNAThr两段基因序列片段。基因定位和序列分析发现,太湖新银鱼线粒体COII基因全序列长度为691 bp,序列AT含量为52.80%,编码230个氨基酸;线粒体Cytb基因序列全长为1141 bp,AT含量为48.90%,它编码380个氨基酸。分别位于线粒体COII和Cytb基因两翼的4个tRNA基因(tRNAAsp、tRNALys、tRNAGlu和tRNAThr)同时被测定出来。将太湖新银鱼与有明银鱼、小齿日本银鱼的同源序列进行比对分析,并基于线粒体COII+Cytb基因合并数据的核苷酸和氨基酸两种序列形式,以黑斑蛙为外群,对10种鱼类进行分子系统树的构建,结果一致表明:小齿日本银鱼与有明银鱼的亲缘关系近于太湖新银鱼;鲱科与鲑科的亲缘关系近于银鱼科鱼类;此外在本研究硬骨鱼类的4个科中,白鲟科作为原始而古老的类群,是在系统进化的过程中首先分化出来的一支。The complete sequences of mitochondrial COⅡ and Cytb genes and their connected tRNA genes （ tRNA^Asp, tRNA^Lys, tRNA^Glu and tRNA^Thr ） were amplified and determined for N. taihuensis by PCR with five pairs of specific primers. The region tRNA^Asp-coⅡ-tRNA^Lys of N. taihuensis was 863 bp in length containing the 691 -base COIIgene, tRNA^Asp gene （73 bp） and tRNA^Lys genes （75 bp）, and the region tRNA^Ghr-Cytb-tRNA^Thr including the Cytb gene （ 1 141 bp）, the tRNAglu gene （69 bp） and the tRNATM gene of 72 nt. Two segments of N. taihuensis including the mitochondrial COIIand Cytb genes have been deposited in GenBank under the accession number EF151003 and EF490211, respectively. The authors aligned the COIIand Cytb genes sequences of N. taihuensis with S. ariakensis and S. microdon （ the published sequences in GenBank, NC_006918 and NC_004599, respectively） using Clustal X software program. With comparison compared among three sequences of Salangidae, the divergences and base substitutions were analyzed with the MEGA3.1 software. To further confirm the phylogenetic relationships among 3 species of Salangidae, another 7 mt genomes of fishes （S. salar, U12143; O. mykiss, DQ288271; D. cepedianum, NC_008107; S. melanostictus, NC_002616; P. gladius, AY571339; R. porosa, NC_007173 and H.francisci, AJ310141 ）were retrieved from GenBank, R. nigromaculata （ AB043889 ） being used as the outgroup species. DNA molecular phylogenies were derived using MEGA3.1, with neighbor-joining analyses, based on two data sets combining COIIwith Cytb genes sequences at the nueleotides and amino acid level. The phylogenetie analyses results showed that two trees of NJ both strongly supported a close relationship between S. ariakensis and S. mierodon and favored a Salmonidae ＋ Clupeidae elade with high bootstap support values. In addition, our results also suggested that Polyodontidae was the most ancestral lineage among 4 families of Aetinopterygii.

关 键 词：太湖新银鱼 线粒体COⅡ基因 CYTB基因 TRNA基因 

分 类 号：Q785[生物学—分子生物学]