| IRM-2小鼠全长cDNA文库的构建及鉴定 |
| |
| 引用本文： | 王芹,李进,宋力,刘强,岳井银,穆传杰,唐卫生,樊飞跃. IRM-2小鼠全长cDNA文库的构建及鉴定[J]. 中华放射医学与防护杂志, 2010, 30(1): 274-278. DOI: 10.3760/cma.j.issn.0254-5098.2010.03.011 |
| |
| 作者姓名： | 王芹 李进 宋力 刘强 岳井银 穆传杰 唐卫生 樊飞跃 |
| |
| 作者单位： | 中国医学科学院放射医学研究所天津市分子核医学重点实验室,天津,300192; |
| |
| 摘 要： | Objective To screen and isolate the radioresistance related genes of IRM-2 mice.Methods cDNA library of IRM-2 mice was constructed by SMART technique.Total RNA was isolated from spleens of IRM-2 male mice.The first-strand cDNA was synthesized by using PowerScript reverse transeriptase,and double-strand cDNA was synthesized and amplified by long PCR.The PCR products were purified,digested with restriction enzyme Sfi I.The ds-cDNA fragment lessthan 500 bp was fractionated and ligated to the Sfi I-digested pDNR-LIB vector.The ligation mixture was transformed into E.coil DH5α by electroporution transformation to generate the unamplified cDNA library.The quality of cDNA library was identified by PCR technique.130 clones from cDNA library were sequenced and compared with GenBank database.Results The cDNA library contained 2.25 x 106 independent clones with an average insert size of 1.2 kb.The ratio of recombination and full-length was 95% and 55%,respectively.21 pieces of EST sequences from cDNA library were not the same as the known mice genes and registered into GenBank EST database,with registered number DW474856-DW474876.Conclusions cDNA library of IRM-2 mice has been constructed successfully.21 pieces of EST implies that radioresistance correlative genes may be in IRM-2 mice,which will lay a foundation for isolating and identifying radioresistance related genes in further study.
|
| 关 键 词： | IRM-2小鼠 cDNA文库 SMART技术 |
Construction and characterization of cDNA library for IRM-2 mice |
| |
| WANG Qin,LI Jin,SONG Li,LIU Qiang,YUE Jing-yin,MU Chuan-jie,TANG Wei-sheng,FAN Fei-yue. Construction and characterization of cDNA library for IRM-2 mice[J]. Chinese Journal of Radiological Medicine and Protection, 2010, 30(1): 274-278. DOI: 10.3760/cma.j.issn.0254-5098.2010.03.011 |
| |
| Authors: | WANG Qin LI Jin SONG Li LIU Qiang YUE Jing-yin MU Chuan-jie TANG Wei-sheng FAN Fei-yue |
| |
| Abstract: | |
| |
| Keywords: | IRM-2 micecDNA librarySMART techique |
|
|
