| WANG Fengchao,WANG Junping,SU Yongping.Cloning of radiation-induced new gene RS1 expressed in mouse intestinal epithelium by enhanced RACE[J].Chinese Journal of Radiological Medicine and Protection,2003,23(5):324-326 |
| Cloning of radiation-induced new gene RS1 expressed in mouse intestinal epithelium by enhanced RACE |
| Received:April 10, 2003 |
| DOI： |
| KeyWords:Enhanced RACE PCR RT-PCR Probe hybridization RS1 gene |
| FundProject:国家自然科学基金资助项目(39970232,30230360) |
| Author Name | Affiliation | | WANG Fengchao | The Institute of Combined Injury, Third Military Medical University, Chongqing 400038, China | | WANG Junping | The Institute of Combined Injury, Third Military Medical University, Chongqing 400038, China | | SU Yongping | The Institute of Combined Injury, Third Military Medical University, Chongqing 400038, China | | 高京生 | The Institute of Combined Injury, Third Military Medical University, Chongqing 400038, China | | 楼淑芬 | The Institute of Combined Injury, Third Military Medical University, Chongqing 400038, China | | 刘晓宏 | The Institute of Combined Injury, Third Military Medical University, Chongqing 400038, China | | 任泂 | The Institute of Combined Injury, Third Military Medical University, Chongqing 400038, China | | 章波 | The Institute of Combined Injury, Third Military Medical University, Chongqing 400038, China |
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| Abstract:: |
| Objective To obtain full-length cDNA of radiation-induced new gene RS1 expressed in mouse intestinal epithelium. Methods The tissue expression profile of RS1 was analyzed by semi-quantitative RT-PCR to find the target tissue which highly expresses RS1.The total RNA extracted from the corresponding tissue was taken as the template for reverse-transcription.Enhanced RACE PCR was used to clone the full-length cDNA of RS1,including enrichment of the target gene through biotin-labeled probe for magnetic bead purification and nested PCR. Results About a 2 kb long 3′end was successfully cloned and cloning of the 5′end proceeded well. Conclusion The result is consistent with our experiment design.The set of combined techniques has been identified with the cloning of full-length cDNA from EST sequence especially when the optimal gene-specific primers are not available or the expression level of target gene is low. |
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