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20 2 Vol. 20, No. 2 2002 5 J. Radiat. Res. Radiat. Process. May 2002 DNA 吴 玮 屈昌民 吴代民 ( 0 100101) ( WRE) DNA 60 , Co WRE , , WRE , PMH16 WRE DNA , WRE G418 69. 75% g t , g t ; 2Gy4Gy WRE 36.05% 21. 50% g t , WRE DNA , , , WRE DNA DNA , , WRE , Q782 , R818.74 [ 1] , ( Waldeyer s Ring Equivalent WRE) , DNA , , DNA , WRE , , DNA WRE DNA , ? , DNA , WRE DNA 1 1. 1 , , 20g 2g, 1. 2 60 - 1 Co , 1. 5Gy!min 1. 3 DNA PMH 16, , ( neo g t) A : DMEM+ 10% , B : A + 2( g/ mL PHA+ 100/ mL IL- 2, C : B + 0. 75mg/ mL G418, D : C + 10% XHATM , XHAT M 10g/ mL 15g / mL 0. 2g / mL 5g / mL 25g/ mL DNA ( Promega ) PMH 16 EcoRV , DNA 1. 4 ( PI) , 2Gy4Gy 8h WRE , ( 299004) : , , 1963 8 , 1998 , , : 2001- 08- 15, 2001- 11- 28 2 : DNA 147 , 200 , , B , 5% CO 37 ∀ 2 24h150 # g , PI, 30min, ( Coulter , EPICS- XL , ) , , ( PCD) WRE 1 # 7 10 5mL, 60Gy 1. 5 - DNA : Li ofectamine 5L 1640 95L 30min; PMH16 DNA2g( 100L) , , 6 6 , 2# 10 / mL, 1# 10 / mL , 60 mm , - DNA , 700L 1640 1mL, 37 ∀ , 5hB 48h 1. 6 Davis[ 2] , 200V, 1080F, 1ms , 3∃ 5 , 48h 1. 7 DNA PMH16 neo , ( G418) ; g t - , XHATM , g t EcoRV , DNA , , PMH 16 DNA , B 48h, 96 , 4 4 4 : 1# 10 / mL2# 10 / mL, 1# 10 / mL , C 15∃ 20d, G4 18 g t D , 15d, 3∃ 4d , XHAT M ( ) ,