| 李建祥,傅春玲,陈锐,聂继华,周建伟,童建.氡染毒小鼠骨髓细胞差异表达cDNA文库的构建和初步鉴定[J].中华放射医学与防护杂志,2008,28(1):21-23 |
| 氡染毒小鼠骨髓细胞差异表达cDNA文库的构建和初步鉴定 |
| Construction and identification of subtracted cDNA library in bone marrow cells of radon-exposed mice |
| 投稿时间：2006-12-22 |
| DOI： |
| 中文关键词: 小鼠 氡染毒 抑制性消减杂交 cDNA消减文库 |
| 英文关键词:Mouse Radon exposure Suppression subtractive hybridization Subtracted cDNA library |
| 基金项目:国家自然科学基金资助项目(30371226);教育部博士点专项基金资助项目(20050285009);江苏省重点实验室开放课题(KJS0622) |
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| 中文摘要: |
| 目的 构建氡染毒小鼠骨髓细胞差异表达基因的cDNA文库。方法 采用HD-3型多功能氡室对雄性BALB/c小鼠动态吸入染毒,建立实验动物模型。按照实验动物吸入氡及其子体的累积剂量分为实验组(105 WLM)和对照组(室内本底浓度,1 WLM)。取骨髓细胞提取总RNA,采用SMART和抑制性消减杂交技术,构建正、反向消减cDNA文库,消减cDNA片段插入pMD18-T载体转化大肠杆菌,以含差异表达cDNA片段的菌液为模板进行巢式PCR扩增。结果 获得了完整无降解的总RNA,得到高效率的消减cDNA文库;克隆了244个有插入片段和长度在100～1100bp之间的单克隆。结论 成功构建了氡暴露小鼠骨髓细胞差异表达的正、反向cDNA消减文库。 |
| 英文摘要: |
| Objective To construct and identify subtracted cDNA library in bone marrow cells of mice exposed to radon inhalation. Methods Adult male BALB/c mice, weighing 18-22 g, were placed in a multi-functional radon chamber. One group of mice was exposed to radon up to the accumulative dose of 105 work level month (WLM). The control group of mice was housed in a room with an accumulative dose of 1 WLM. To construct a subtracted cDNA library enriched with differentially expressed genes, the SMART technique and the suppression subtractive hybridization were performed. The obtained forward and reverse cDNA fragments were directly inserted into pMD18-T vector and transformed into E.coli JM109. The inserting cDNA fragments were screened by the blue-and-white blot screening and nested PCR of bacterium liquid.Results The 244 of 285 white bacteria clones obtained randomly were positive clones contained 100-1100 bp inserted cDNA fragments.Conclusions The forward and reverse subtracted cDNA library in bone marrow cells of mice exposed to radon inhalation is successfully constructed. |
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