| RAO Yalan CHEN Xiaohua,ZHANG Junquan,et al.Expressed sequence tags isolated from mouse bone marrow cells irradiated with high-dose gamma rays by suppression subtractive hybridization (SSH)[J].Chinese Journal of Radiological Medicine and Protection,2003,23(3):163-166 |
| Expressed sequence tags isolated from mouse bone marrow cells irradiated with high-dose gamma rays by suppression subtractive hybridization (SSH) |
| Received:September 27, 2002 |
| DOI： |
| KeyWords:Mouse bone marrow Ionizing radiation Suppression subtractive hybridization Expressed sequence tags |
| FundProject:全军“十五”医药卫生科研基金资助项目(01L018) |
| Author Name | Affiliation | | RAO Yalan CHEN Xiaohua | Beijing Institute of Radiation Medicine, Beijing 100850, China | | ZHANG Junquan | Beijing Institute of Radiation Medicine, Beijing 100850, China | | 张军权 | Beijing Institute of Radiation Medicine, Beijing 100850, China | | 魏巍 | Beijing Institute of Radiation Medicine, Beijing 100850, China | | 董波 | Beijing Institute of Radiation Medicine, Beijing 100850, China | | 毛秉智 | Beijing Institute of Radiation Medicine, Beijing 100850, China |
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| Abstract:: |
| Objective To explore the molecular mechanism of bone marrow injury caused by irradiation, gene expression of bone marrow cells in mice after whole body irradiation was studied. Methods Total RNA extracted from bone marrow cells in mice at 4 h after whole body irradiation with 7 Gy γ-rays was taken as the tester and the total RNA from control mice as the driver cDNA subtraction was performed using the protocols described in the Clontech SMART PCR cDNA Sythesis Kit and PCR-select cDNA Subtraction Kit.The subtracted cDNA was then inserted into T vector to generate subtracted cDNA library.Clones of the subtracted cDNA library were screened by hybridization and the insert sequence of the positive clones was compared with the sequence in the GenBank. Results A total of 800 clones selected from the subtracted cDNA library were PCR-amplified and about 86% had inserts.Fourteen differential cDNA fragments were acquired after two round hybridization screening, 7 of them showed high similarity to the expressed sequence tags (ESTs) in the mouse EST database and the other 7 cDNA fragments were possibly new ESTs. Conclusion The succesessfully constructed subtracted cDNA library of bone marrow cells in mice after whole body high-dose irradiation and the validation of some differential ESTs establish the basis for further research of radiation-related genes. |
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