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黑线仓鼠白化突变系抑制差减文库的构建及初步分析

     

摘要

目的 构建黑线仓鼠白化突变系的抑制差减文库,分离获得黑线仓鼠发生被毛突变的相关基因片段,并进行初步分析.方法 以黑线仓鼠作为实验组(tester),以其白化突变系作为驱动组(driver),采用抑制差减杂交技术构建差减文库(SSH).对阳性克隆进行测序并分析.结果 共获得126个有生物学意义的表达序列标签(ESTs),包括21个骨架蛋白,42个转录因子,24个代谢酶基因,25个转运分子及其调节蛋白和4个未知基因.差异基因中没有发现常见的白化疾病相关基因,发现多种与溶酶体的形成及内吞作用相关的基因,推测黑线仓鼠白化突变性状的产生可能与黑色素的运输障碍有关.结论 成功构建了黑线仓鼠白化突变的抑制差减文库,获得126个白化突变的相关基因,为进一步寻找白化突变相关基因,揭示其分子机制奠定了基础.%Objective To construct the subtractive library for Cricetulus barabensis and its albino mutant and preliminarily analyse the obtained gene fragments that related to the hair mutation of Cricetulus barabensis. Methods The Cricetulus barabensis was used as tester and its albino mutant was used as driver, the subtractive Library was constructed by suppression subtractive hybridization (SSH) method. The positive clones was sequenced and analyzed. Results 126 Expressed sequence tags (ESTs) was obtained including 21 skeleton proteins, 42 transcription factors, 24 genes of metabolic enzymes, 25 transport factors and their regulators, and 4 genes of unknown. Not the common genes related to albinism but several kinds of genes related to the formation and endocytosis of lysosomes were found in the differentially expressed genes, suggesting that the production of albinism of Cricetulus barabensis may relate to the disfunction of transportation. Conclusion The subtractive library for Cricetulus barabensis and its albino mutant was successfully constructed and 126 ESTs was obtained. This study may lay a basis for looking for the genes related to albinism of Cricetulus barabensis and revealing the corresponding mechanism.

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