| 乌克兰鳞鲤同工酶及mtDNA D-loop区段的RFLP分析 |
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| 引用本文： | 梁爱军,王淞,鲍迪,孙少起,董仕.乌克兰鳞鲤同工酶及mtDNA D-loop区段的RFLP分析[J].安徽农业科学,2011,39(27):17109-17113. |
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| 作者姓名： | 梁爱军 王淞 鲍迪 孙少起 董仕 |
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| 作者单位： | 1. 天津师范大学生命科学学院,天津市细胞遗传与分子调控重点实验室,天津300387
2. 天津市凯润淡水养殖有限公司,天津,300385 |
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| 基金项目： | 天津师范大学校内基金项目(5RL021) |
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| 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。
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| 关 键 词： | 乌克兰鳞鲤 同工酶分析 mtDNA D-loop RFLP DNA测序 |
Analysis of Isozyme and MtDNA D-loop Region of Ukraine Scale Carp by RFLP |
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| LIANG Ai-jun et al.Analysis of Isozyme and MtDNA D-loop Region of Ukraine Scale Carp by RFLP[J].Journal of Anhui Agricultural Sciences,2011,39(27):17109-17113. |
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| Authors: | LIANG Ai-jun |
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| Institution: | LIANG Ai-jun et al(College of Life Sciences,Tianjin Normal University,Tianjin Key Laboratory of Cyto-Genetical and Molecular Regulation,Tianjin 300387) |
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| Abstract: | 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. |
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| Keywords: | Ukraine scale carp Isozyme analysis mtDNA D-loop RFLP DNA sequencing |
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