高温致神经管畸形相关基因CDK109的克隆及其致畸相关性分析.docVIP

  高温致神经管畸形相关基因 CDK109 的克 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* 5 10 15 20 25 30 35 40 （1. 山东大学医学院人体解剖学教研室，济南 250012； 2. 山东大学医学院组织胚胎学教研室，济南 250012） 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的神经管 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 文库中筛选出了高温 致畸相关基因 CDK109 全长，其在高温致畸组的表达明显高于正常对照组。 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：神经管畸形；高温；cDNA 文库；CDK109；金黄地鼠 中图分类号：R361.2 Cloning and relativity analysis of the gene-CDK109 related to neural tube defects induced by hyperthermia ZHANG Jing1, GAO Yingmao2, SUN Jinhao1, BAO Lihua1 (1. Department of Human Anatomy, Shandong University School of Medicine,Jinan 250012; 2. Department of Histology and Embryology, Shandong University School of Medicine,Jinan 250012) Abstract: Objective To screen some potential genes from the cDNA library of neural tube from golden hamster treated with hyperthermia, clone these genes and analyze their relativity with the neural tube defects（NTDs） induced by hyperthermia. Methods With Benton-Davis hybridization, a piece of oligonucleotide probe was used to screen the cDNA library of neural tube from golden hamster treated with hyperthermia which had been constructed by our laboratory. Then the positive phage clone was transformed into E. coli BM25.8 and the plasmid was isolated and identified by restriction enzyme digestion and the positive plasmid was sequenced and blasted. By Northern blot, the expression of this screened gene-CDK109 in heat-treated and control groups of the golden hamsters at day 8.5 after mating was detected and reconfirmed. Results The whole length of CDK109 gene was successfully cloned from cDNA library of neural tube from golden hamster treated with hyperthermia. Its expression in heat-treated groups was evidently higher than control groups. Conclusion The increasing expression of CDK109 is intimately related with neura