摘要: |
于1996年4月在中国科学院水生生物研究所关桥实验场取得银鲫E系(8♀,2♂)和D系(9♀,1♂)的卵巢(♀)或肝脏(♂)组织,分离纯化其线粒体DNA,以此为材料,用25种限制性内切酶分析了银鲫这两个雌核发育系的线粒体DNA,构建了两系鱼mtDNA的19种酶41个位点的酶切图谱。统计比较酶切片段的大小,发现5种内切酶(AvαⅡ, BamHⅠ, BglⅠ, SphⅠ, XbaⅠ)酶切位点在两个雌核发育系间存在差异。这些多态性,既可以作为区分两个雌核发育系的遗传标记,同时也为其进化遗传学的研究和银鲫育种提供了丰富的资料。 |
关键词: 雌核发育银鲫 限制性内切酶 mtDNA 遗传标记 |
DOI:10.11693/hyhz200004004004 |
分类号: |
基金项目:国家杰出青年科学基金资助项目,39425010号 |
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COMPARATIVE STUDIES ON MITOCHONDRIAL DNAs OF TWO DIFFERENT CLONES OF CARASSIUS AURATUS GIBELIO |
FAN Lian-chun,LAI Yu-peng,ZHU Lan-fei,LIANG Shao-chang,GUI Jian-fang
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State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, The Chinese Academy of Sciences, Wuhan430072
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Abstract: |
Oocytes (female) and livers (male) from E clone (8 female, 2 male) and D clone (9 female, 1 male) of silver crucian carp were collected from Guanqiao experiment center in Institute of Hydrobiology of Chinese Academy of Sciences in April 1996. The mitochondrial DNAs from the two different clones (E and D clones) of gynogenetic silver crucian carp were isolated and digested with 25 restriction endonuleases: ApαI,, AvaⅡ, BamHI, BglI, BglⅡ, BstEⅡ, BstXI, EcoRI, EcoRV, HindIII, HinfI, KpnI, MspI, NcoI, NsiI, PstI, PvuⅡ, SacI, SalI , SmaI , SphI , SspI, StyI, XbaI, XhoI. physical maps of the restriction endonucleases cleavage sites on the mtDNAs of the two clones of silver crucian carp were drawn according to the numbers and sizes of the fragments by single and double digestion, which include total 41 cleavage sites from 19 endonucleases. All of the endonucleases produce 1–10 fragments except XhoI which has no cut site in both fish mtDNAs. Comparing the digestion results, we found 5 restriction endonucleases (AvaⅡ, BamHI, BglI, SphI, XbaI) with cleavage site differences. These differences can be used as the genetic marker for the two different clones of silver crucian carp in genetic breeding practice. Using the results of the endonucleases digestion, the homologous extent between the two fish clones can be calculated. The basic substitution rate of the two fish clone is P=0.010118, which shows the divergence date of the two fish clones is probably 267 thousand years or 165 thousand years age. |
Key words: Gynogenetic Carassius auratus gibelio, Restriction endonuclease, mtDNA, Genetic marker |