黑线仓鼠白化突变系抑制差减文库的构建及初步分析

doi:10.3969/j.issn.1674-5817.2013.01.004

实验动物与比较医学 ›› 2013, Vol. 33 ›› Issue (1): 19-22.DOI: 10.3969/j.issn.1674-5817.2013.01.004

黑线仓鼠白化突变系抑制差减文库的构建及初步分析

李爱学, 曾林, 尚世臣, 王鹏, 姚晓兰, 孙兆增

军事医学科学院实验动物中心, 北京100071

收稿日期:2012-07-17 出版日期:2013-02-25 发布日期:2013-02-25
作者简介:李爱学(1978-), 女, 博士, 研究方向: 实验动物学。E-mail: lax200178@yahoo.com.cn。
基金资助:
国家自然基金(31272387、31071987), 军事医学科学院实验动物中心青年基金项目(QN2010-003)

Construction and Preliminary Analysis of Subtractive Library for Cricetulus Barabensis with Albino Mutant

LI Ai-xue, ZENG Lin, SHANG Shi-chen, WANG Peng, YAO Xiao-lan, SUN Zhao-zeng

Laboratory Animal Center of the Academy of Military Medical Science, Beijing 100071, China

Received:2012-07-17 Online:2013-02-25 Published:2013-02-25

摘要/Abstract

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。

关键词: 黑线仓鼠, 白化突变, 差减杂交, 筛选

Abstract: 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.

Key words: Cricetulus barabensis, Albino mutant, SSH, Construction

中图分类号:

Q95-33

李爱学, 曾林, 尚世臣, 王鹏, 姚晓兰, 孙兆增. 黑线仓鼠白化突变系抑制差减文库的构建及初步分析[J]. 实验动物与比较医学, 2013, 33(1): 19-22.

LI Ai-xue, ZENG Lin, SHANG Shi-chen, WANG Peng, YAO Xiao-lan, SUN Zhao-zeng. Construction and Preliminary Analysis of Subtractive Library for Cricetulus Barabensis with Albino Mutant[J]. Laboratory Animal and Comparative Medicine, 2013, 33(1): 19-22.

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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

Construction and Preliminary Analysis of Subtractive Library for Cricetulus Barabensis with Albino Mutant

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