| 摘要: |
| 采用FIASCO (Fast Isolation by AFLP of Sequences Containing repeats)法和标记初步筛选法, 进行了大菱鲆(Scophthalmus maximus)微卫星标记开发的研究, 分析了大菱鲆基因组中微卫星DNA序列的特征, 并进行了拟作图标记的初步筛选。结果表明, FIASCO法的富集效率为78.56%, 二次筛选出655个阳性克隆, 测序后经分析有469条含有微卫星位点的单一序列, 共得到597个微卫星位点。其中完美型占51.76%, 非完美型占34.67%, 复合型占13.57%; 核心序列的重复次数在3—96次之间, 平均重复数为13.39次; 重复单元类别最多的是(CA/GT)n, 占77.6%。利用Primer Premier 5.0共设计413对引物, 其中有360对能够得到稳定表达的产物; 再经标记初筛试验得知, 183对引物(50.8%)的PCR产物有多态性, 其中110个微卫星位点(30.6%)符合遗传连锁图谱作图标准。 |
| 关键词: 大菱鲆 微卫星标记 标记开发 序列分析 标记初筛 |
| DOI：10.11693/hyhz201305036036 |
| 分类号: |
| 基金项目:现代农业产业技术体系建设专项资金资助, CARS-50-G01号。 |
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| ISOLATION AND ANALYSIS OF MICROSATALLITE MARKERS IN THE GENOME OF TURBOT SCOPHTHALMUS MAXIMUS |
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LI Meng1, MA Ai-Jun2, YUE Liang2, ZOU Jie2, WANG Guang-Ning2, TIAN Yue-Qiang2, MA Ben-He2, XIA Dan-Dan2
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1.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences,Fisheries and Life Science, Shanghai Ocean University;2.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences
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| Abstract: |
| We constructed microsatellite markers of turbot Scophthalmus maximus by FIASCO (Fast Isolation by AFLP of Sequences Containing repeats) method to analyze the microsatellite DNA sequences of the turbot genome, applying the FIASCO method for initial screening of the markers plotted. The results indicate that enrichment efficiency by the method FIASCO method is 78.56%. Among 655 positive clones secondly screened, 597 were microsatellite loci and 469 were unique sequences. Of the 597 microsatellite loci, 51.76% were perfect, 34.67% imperfect, and 13.57% compound. The core sequence repetitions ranged 3—96 times; the average number of duplicates was 13.39; the most repeat unit type was the (CA/GT)n, 77.6%. With Primer Premier 5.0, 413 pairs of primers were designed, of which 360 could be stably expressed. In addition, using marker screening again, 183 pairs of primers (50.8%) was found polymorphism in PCR product, from which 110 microsatellite loci (30.6%) were identified in compliance to the standards of genetic linkage mapping. |
| Key words: turbot Scophthalmus maximus microsatellite markers marker isolation sequence analysis preliminary screening |
