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(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,成功构建了氡暴露小鼠外周血细胞差异表达的cDNA消减文库。
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.
出处
《科学技术与工程》
2007年第3期273-276,共4页
Science Technology and Engineering
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
