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山东大学学报(医学版) ›› 2012, Vol. 50 ›› Issue (1): 19-.

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高温致神经管畸形相关基因CDK109克隆及其致畸相关性分析

张静1,高英茂2,孙晋浩1,暴丽华1   

  1. 山东大学医学院1.人体解剖学教研室;2.组织学与胚胎学教研室,济南   250012
  • 收稿日期:2011-04-12 出版日期:2012-01-10 发布日期:2012-01-10
  • 作者简介:张静(1978- ),女,博士,讲师,主要从事实验畸形学的研究。
  • 基金资助:

    教育部高等学校博士学科点专项科研基金新教师类资助项目(20090131120031), 山东省自然科学基金青年基金资助项目(ZR2009CQ010)

Cloning and relativity analysis of gene CDK109 related to neural tube defects induced by hyperthermia

ZHANG Jing1, GAO Ying-mao2, SUN Jin-hao1, BAO Li-hua1   

  1. 1. Department of Human Anatomy; 2. Department of Histology and Embryology,
     School of Medicine, Shandong University, Jinan 250012, China
  • Received:2011-04-12 Online:2012-01-10 Published:2012-01-10

摘要:

目的   通过筛选高温致畸金黄地鼠胚胎神经管cDNA文库,克隆高温致畸相关基因CDK109。方法   利用本室构建的高温致畸胚胎神经管cDNA文库,采用噬菌斑原位杂交方法,用寡核苷酸探针筛选cDNA文库,挑取阳性噬菌斑,将其转化为质粒,再通过限制性酶切鉴定阳性克隆并测序,将该基因标记探针与孕8.5d高温致畸胚胎神经管组织和正常对照胚胎神经管组织总RNA进行NORTHERN杂交,以便确认该基因在高温致神经管畸形中的异常表达。 结果   成功地从高温致畸金黄地鼠胚胎神经管cDNA文库中筛选出高温致畸相关基因CDK109全长,其在高温致畸组的表达明显高于对照组。 结论   CDK109在神经管中的高表达与高温致神经管畸形密切相关。

关键词: 神经管缺损;高温,诱发;cDNA文库;基因,CDK109;金仓鼠

Abstract:

Objective   To screen for gene CDK109 from the cDNA library of the neural tube from golden hamster treated with hyperthermia, clone this gene and analyze its relativity with neural tube defects(NTDs) induced by hyperthermia. Methods   With BentonDavis hybridization, a oligonucleotide probe was used to screen the cDNA library of the neural tube from golden hamster model treated with hyperthermia, which had been constructed in our laboratory. Then the positive phage clone was transfected into E.coli BM25.8, the plasmid was isolated and identified by restriction enzyme digestion, and the positive plasmid was sequenced and blasted. By Northern blot, expression of screened gene CDK109 was detected and reconfirmed in heat-treated and control groups of golden hamsters on pregnant day 8.5. Results   The whole length of the CDK109 gene was successfully screened from the cDNA library of the neural tube from golden hamsters treated with hyperthermia. Its expression in the heat-treated group was evidently higher than in the control group.  Conclusion   Increased expression of CDK109 is closely related to neural tube defects induced by hyperthermia.

Key words: Neural tube defects; Hyperthermia, induced; cDNA library; Genes, CDK109; Mesocricetus

中图分类号: 

  • R361.2
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