氡染毒小鼠肺及支气管组织的基因表达  

Experimental study of gene expression in lung and bronchus of radon-exposed mice

作　　者：郭志英[1] 田梅[1] 刘建香[1] 阮健磊[1] 朴春南[1] 苏旭[1] 

机构地区：[1]中国疾病预防控制中心辐射防护与核安全医学所,北京100088

出　　处：《中华放射医学与防护杂志》2008年第4期335-338,共4页Chinese Journal of Radiological Medicine and Protection

摘　　要：目的 构建氡染毒小鼠肺及支气管组织差异表达基因的cDNA文库，并对其进行初步鉴定和同源性分析。方法 采用SR—NIM02型氡室对小鼠进行吸人染毒后，常规饲养3个月，分别提取和纯化染毒组（30工作水平月，WI。M）与对照组（0．02WLM）小鼠的肺及支气管组织的总RNA，采用Super SMART技术和抑制性差减杂交技术（SSH），构建氡染毒后肺及支气管组织的正、反向差减杂交的cDNA文库；常规连接pGEM—T—easv载体，转化DH5α感受态细胞，巢式PCR鉴定；对阳性克隆测序，并与GeneBank数据库进行BLAST同源性比对，进行初步功能分类。结果共获得克隆460个，其中含有插入片段的克隆146个，经BLAST同源性比对后有48个正向差减cDNA片段与61个反向差减cDNA片段与GeneBank中的序列有不同程度的同源性。结论 成功构建了氡染毒小鼠肺支气管差异表达cDNA文库，染毒后小鼠肺及支气管组织中某些基因会发生差异表达，其中部分基因可能参与细胞凋亡和周期调控、机体免疫调节及细胞间信号传导等过程。Objective To construct and identify differentially expressed cDNA library in lung and bronchus of mice exposed to radon. Methods 2 week old, weighing （18-22）g, male BALB/c mice were placed in a SR-NIM02 radon chamber. One group of mice was exposed to radon, which was equivalent to the accumulative dose of 30 WLM. The control group was about 0.02 WLM. To construct a subtracted cDNA library enriched with differentially expressed genes, the Super SMART technique and the suppression subtractive hybridization （SSH） were performed. The obtained forward and reverse cDNA fragments were directly inserted into pGEM-T-easy vector and transformed into E. coli DH5α. The inserts in plasmid were amplified by nested polymerase chain reaction （PCR）, and some of which were sequenced. In the end these sequences were BLASTed with GeneBank. Results 146 of 460 clones obtained randomly were positive clones contained （1000- 1500）bp inserted cDNA fragments. The forward and reverse subtracted cDNA library in lung and bronchus of mice exposed to radon was constructed, and 48 up-regulation and 61 down-regulation cDNA sequences selected were homologous with GeneBank in different extent. Conclusions The subtracted cDNA library in lung and bronchus of mice exposed to radon is successfully constructed, and genes that differentially expressed are identified. Some genes might have relation with the immunity, cell cycle and apoptosis.

关 键 词：小鼠 氡及其子体 染毒 抑制差减杂交 cDNA差减文库 

分 类 号：R114[医药卫生—卫生毒理学]