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氡染毒小鼠外周血细胞差异表达cDNA文库的构建和初步鉴定
  • ISSN号:1671-1815
  • 期刊名称:科学技术与工程
  • 时间:0
  • 页码:487-499
  • 语言:中文
  • 分类:Q691.9[生物学—生物物理学] Q78[生物学—分子生物学]
  • 作者机构:[1]苏州大学放射医学与公共卫生学院卫生毒理学教研室, [2]苏州大学放射医学与公共卫生学院营养与食品教研室,苏州215123, [3]苏州大学附属第二医院呼吸内科,苏州215004
  • 相关基金:国家自然科学基金(30371226),教育部博士学科点专项科研基金(20050285009)资助.
  • 相关项目:吸烟与氡暴露的蛋白质组差异表达及肺癌早期蛋白标志物的筛选研究
中文摘要:

构建氡染毒小鼠外周血细胞差异表达基因的cDNA文库。采用HD-3型多功能氧室对雄性BALB/C小鼠动态吸入染毒,建立实验动物模型。按照实验动物吸入氧及其子体的累积荆量分为实验组(105WLM)和对照组(室内本底浓度,1WLM)。提取外周血细胞总RNA,采用SMART和抑制性消减杂交技术,构建正、反向消减cDNA文库,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。获得了312个单克隆,成功构建了氧暴露小鼠外周血细胞差异表达的cDNA消减文库。

英文摘要:

This study was designed to construct and identify subtracted cDNA library in peripheral ceils of mice exposed to radon inhalation. Adult male BALB/c mice weighing ( 18 - 22) g were placed in a muhi-functional radon chamber. One group of mice was exposed to radon, 12 h per day, which was equivalent to the accumulative dose of 105WLM. The control group of mice was housed in a room with a accumulative dose of 1 WLM. To construct a subtracted cDNA library enriched with differentially expressed genes, the SMART technique and the suppression subtractive hybridization was performed. The obtained forward and reverse cDNA fragments were directly inserted into pMD - 18 vector and transformed into E. coli JM109. The inserting cDNA fragments were screened by blue and while blot screening and nested PCR of bacterium liquid. The 312 of 390 white bacteria clones obtained randomly were positive clones contained ( 100 - 1 000) bp inserted cDNA fragments. The forward and reverse subtracted cDNA library in peripheral blood cells of mice exposed to radon inhalation was successfully constructed.

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期刊信息
  • 《科学技术与工程》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学技术协会
  • 主办单位:中国技术经济学会
  • 主编:明廷华
  • 地址:北京市学院南路86号
  • 邮编:100081
  • 邮箱:ste@periodicals.net.cn
  • 电话:010-62118920
  • 国际标准刊号:ISSN:1671-1815
  • 国内统一刊号:ISSN:11-4688/T
  • 邮发代号:2-734
  • 获奖情况:
  • 国内外数据库收录:
  • 中国中国科技核心期刊,中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:29478