| LI Jian-xiang,FU Chun-ling,CHEN Rui,et al.Construction and identification of subtracted cDNA library in bone marrow cells of radon-exposed mice[J].Chinese Journal of Radiological Medicine and Protection,2008,28(1):21-23 |
| Construction and identification of subtracted cDNA library in bone marrow cells of radon-exposed mice |
| Received:December 22, 2006 |
| DOI： |
| KeyWords:Mouse Radon exposure Suppression subtractive hybridization Subtracted cDNA library |
| FundProject: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) |
| Author Name | Affiliation | E-mail | | LI Jian-xiang | Department of Hygienic Toxicology, School of Radiation Medicine and Public Health, Soochow University, Suzhou 215123, China | | | FU Chun-ling | 营养与食品教研室 | | | CHEN Rui | 苏州大学附属第二医院呼吸内科 | | | 聂继华 | Department of Hygienic Toxicology, School of Radiation Medicine and Public Health, Soochow University, Suzhou 215123, China | | | 周建伟 | 南京医科大学 | | | 童建 | Department of Hygienic Toxicology, School of Radiation Medicine and Public Health, Soochow University, Suzhou 215123, China | tongjian@suda.edu.cn |
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| Abstract:: |
| 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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