文献类型：期刊文献

中文题名：5种笛鲷mtDNA及Cytb基因片段的RFLP比较

英文题名：Comparison on RFLP of mitochondrial DNA and Cyt bgene segment for 5 species in Lutjanus

作者：王中铎[1];刘楚吾[1];郭昱嵩[1]

机构：[1]湛江海洋大学水产学院,广东湛江524025

年份：2005

卷号：29

期号：3

起止页码：327

中文期刊名：水产学报

外文期刊名：Journal of Fisheries of China

收录：CSTPCD、、北大核心2004、Scopus、CSCD2011_2012、北大核心、CSCD

基金：广东省科技计划项目重大专项(A3050202);湛江市科技攻关项目(200017)

语种：中文

中文关键词：笛鲷属;线粒体DNA限制性片段长度多态性分析;PCR-RFLP;系统发生;细胞色素b;假基因

外文关键词：Lutjanus; restriction fragment length polymorphism of mitchondrial DNA; polymerase chain reaction; phylogenesis; cytochrome b; Numts

中文摘要：采用线粒体DNA限制性片段长度多态性分析(mtDNARFLP)和线粒体DNA的细胞色素b基因(Cytb)的部分序列扩增限制性片段长度多态性分析(mtDNAPCRRFLP)两种方法,对笛鲷属5种鱼类进行种间系统发育的比较研究,结果显示:(1)画眉笛鲷依照其形态学上体侧条带颜色差异,可分为褐带画眉笛鲷和黄带画眉笛鲷两个种;(2)千年笛鲷和星点笛鲷、褐带画眉笛鲷和黄带画眉笛鲷、金带笛鲷和金焰笛鲷之间遗传距离相对较近;(3)两种分析方法都可以为种的区分提供方便有效的分子标记;(4)两种分析方法在构建发育系统树上不完全一致。本文从mtDNA角度深入研究了南海海域笛鲷的系统发生,表明了mtDNA作为一种广泛应用的分子标记的实用性,同时也有一些潜在问题需要进一步研究。

外文摘要：The phylogenetic relationships of 5 species of snappers (Lutjanus stellatus, L.sebae,L.vitta, L.fulvus and L.fulviflamma) were studied comparatively, which were based on 2 methods: restriction fragment length polymorphism of mitchondrial DNA (mtDNA-RFLP), RFLP of cytochrome b (Cyt b) gene segment(>900bp) amplified by polymerase chain reaction(PCR-RFLP). The sample numbers of Lutjanus stellatus, L.sebae, L.vitta, L.fulvus and L.fulviflamma were 17,17,35,17 and 17, respectively, which are collected in the South China Sea from Sep.2002 to Jun.2003. The mtDNA being isolated and purified from fresh livers of samples range from 17.05±0.46 to 17.77±0.91 kb. Having been digested by 19 restriction endonucleases recognized 6 base pairs(EcoRⅠ, HindⅢ, BglⅠ, BglⅡ, StyⅠ, XbaⅠ, DraⅠ, PstⅠ, PvuⅡ, XhoⅠ, BamHⅠ, ScaⅠ, EcoRⅤ,SmaⅠ, MluⅠ, SalⅠ, HinfⅠ, KpnⅠ,HpaⅠ), the mtDNA were analysed. The result of mtDNA-RFLP shows that L.vitta could be divided into HE and HD, based on the color of stripe on the middle of the side. The physical maps of mtDNA from L.stllatus and L.sebae have been constructed based on the endonucleases that are chosen from 19 restriction endonucleases.By comparison of the sequence of cytochrome b gene PCR-RFLP using different amplification templates (one from purified mtDNA and the other from nDNA), the possible interferences of nuclear mitochondrial pseudogenes (Numts) were excluded and the possible pseudogenes from nDNA were preliminarily discussed. Having been digested by 6 restriction endonucleases recognized 4 base pairs(StyⅠ, HinfⅠ, HpaⅡ, MspⅠ, Sau96Ⅰ, SinⅠ, TaqⅠ, HaeⅡ), the sequences were analysed. The results of mtDNA PCR-RFLP support that L.vitta could be divided into two species: HE and HD by the 0.0751 of divergency index. The results reveal that all of the 2 analysis methods are useful for seeking out the genetic markers among species effectively. And the results from RFLP indicate that the L.vitta could be divided into two species based on the color of stripe on the middle of the side. At the same time, the constructions of phylogenic trees by neighbor-joining(NJ) method show that there are some differences along different methods on the specific level when they are used for phylogenesis.