外源小鼠核酸促电离辐射损伤的同系小鼠肠腺细胞修复的相关基因克隆

CLONING OF MOUSE GENES RELATED TO REPAIRING OF INTESTINAL EPITHELIUM OF THE γ IRRADIATED MICE BY TREATMENT WITH THE INTESTINAL RNA OF MICE OF THE SAME STRAIN

为研究克隆外源核酸促电离辐射损伤的小鼠肠腺细胞修复的相关基因。建立BALB/c小鼠电离辐射后给予外源RNA与生理盐水治疗的 6、 12、 2 4h ,4d和 8d的模型 ,收集空肠组织标本后采用消减杂交基础上的LD -PCR技术 ,获取与受照射小鼠肠腺损伤修复相关的基因克隆 ,对其进行全自动测序与GeneBank检索 ,新基因递交给基因库。获取了 90个与肠腺修复相关的基因 ,杂交证实在核酸治疗组与生理盐水治疗组之间呈差异表达 ,其中 18个是新基因 ,GeneBank接受号为AF2 4 0 164- 2 4 0 181。获取了 90个可能与外源核酸促肠腺修复相关的基因克隆 ,其中

To clone the new genes involved in the repair of radiation-damaged intestinal crypts of mice which were treated with the intestinal RNA of the mice of the same strain. As a test group, 45 mice which had been irradiated by γ rays were injected with intestinal RNA of mice of the same strain in 2h after irradiation and the specimens of the small intestine of these animals were collected at 6,12,24h,4d and 8d after irradiation respectively. The other 45 mice ,as a control group, were treated with irradiation and physiological saline,and the specimens were collected as those in the test group. The genes which expressed more highly in the test group than in the control were cloned into T vectors after subtractive hybridization and LD-PCR and then sequenced. The sequences obtained were aligned through GeneBank for the new gene search. 90 clones were found associate with the repairing of radiation-damaged mouse intestinal crypts,which was confirmed by RNA dot blot assays. Among the 90 clones, 18 were accepted by GeneBank as new genes with the acceptance numbers AF240164-AF240181. Obtained ninety clones may be correlated closely with repairing of intestinal epithelium of the γ -irradiated mice by treatment with the intestinal RNA of mice of the same strain.