摘要
[目的]分析乌克兰鳞鲤的遗传结构,以期为乌克兰鳞鲤的遗传育种研究提供依据。[方法]利用水平式淀粉凝胶电泳法对乌克兰鳞鲤(46尾)进行了AAT、α-GPD、GPI、IDH、MDH和PGM共6种同工酶以及肌浆蛋白(PROT)的电泳分析。应用PCR技术扩增了其mtDNA的D-loop区段,利用12种限制性内切酶对其扩增片段进行了RFLP分析;并对2种单倍型的PCR扩增片段进行了序列分析。[结果]同工酶检测出的12个基因座位中,α-GPD*、GPI*和PGM*存在变异,多态基因座位比例及平均杂合度预期值分别为25%和0.092 0。RFLP检测结果表明PCR扩增到约1.6 kb的DNA片段,8种限制性内切酶(AluⅠ、DraⅠ、EcoRⅠ、HaeⅢ、HapⅡ、HinfⅠ、TaqⅠ和XspⅠ)有酶切位点,DraⅠ和TaqⅠ个体间存在变异。将不同酶切类型结果组合后,得到2种单倍型。[结论]根据同工酶检测结果,可以认为乌克兰鳞鲤含有中国鲤与欧洲鲤2种血统;与RFLP分析结果相比,序列分析表明2种单倍型在DraⅠ和TaqⅠ酶切位点以外也存在碱基差异。
[Objective] The research aimed to analyze the genetic structure of Ukraine Scale Carp and provide basis for its heredity and breeding study.[Method] The horizontal starch gel electrophoresis was used to evaluate the genetic difference in 6 isozymes（AAT,α-GPD,GP,IDH,MDH,and PGM） and general protein（PROT） in 46 Ukraine scale carp.Its mtDNA D-loop region was amplified by PCR and the amplification fragments were analyzed using 12 kinds of restriction endonuclease by RFLP.And PCR amplification fragments of two haplotypes were sequenced.[Result] The results of isozyme suggested that twelve loci were obtained,three loci（α-GPD＊,GPI＊ and PGM＊） of which were polymorphic.The proportion of polymorphic loci was 25% and expected heterozygosity was 0.092 0.The result of RFLP analysis indicated that the amplified fragment length of mtDNA D-loop region was about 1.6 kb,eight restriction endonucleases（Alu Ⅰ,Dra Ⅰ,EcoR Ⅰ,Hae Ⅲ,Hap Ⅱ,Hinf Ⅰ,Taq Ⅰ and Xsp Ⅰ） had restriction enzyme cutting sites and 2 of which（Dra Ⅰ and Taq Ⅰ） were various.[Conclusion] It can be concluded that the Ukraine scale carp is hybrid of Chinese and European common carp according to the isozyme＇s results.The sequence of mtDNA D-loop of two haplotypes obtained more variation out of the restriction endonucleases site,compared with RFLP analysis.
出处
《安徽农业科学》
CAS
北大核心
2011年第27期17109-17113,共5页
Journal of Anhui Agricultural Sciences
基金
天津师范大学校内基金项目(5RL021)
