摘要
用电脉冲介导和DNA-磷酸钙共沉淀两种基因转移方法,将人HeLaS 3细胞DNA及限制性核酸内切酶切割后的DNA片段,与选择标记基因一同导入XP细胞,经两次选择后得到转化的细胞。实验证明HeLaS3 DNA及经Bgl I、Xho I切割后的DNA片段可以修复XP细胞切除修复基因的缺陷,提高它们对紫外辐射的抗性。二次转化实验进一步证明,导入的HeLaS 3 DNA已在XP细胞基因组中整合,并得到稳定的表达。
The DNA-mediated gene transfer had been carried out by bothCalcium phosphate coprecipitation and electroporation method. The cellular DNA andDNA fragments from human cervical carcinoma HeLaS3 cells were introduced withPSV2Neo DNA into XP20S (SV40) cells. The transfectants were picked up aftertwice selections by G418 and 3J/m^2 UV-irradiation. The results showed that cellularDNA and BglⅠ, XhoⅠ digested DNA fragments from HeLaS 3 could correct thedeficiency of excision repair gene in XP cells, and cause the recipient cells resistantto UV irradiation. The second transfection experiment confirmed that HeLaS3 DNAwere really integrated into XP cell chromosome and stably expressed within the cellgenome.
出处
《辐射研究与辐射工艺学报》
CAS
CSCD
北大核心
1989年第4期25-29,共5页
Journal of Radiation Research and Radiation Processing
关键词
紫外辐射
基因转移
XP细胞
DNA
人
UV-irradiation
Excision repair
Gene transfer
XP cell
HeLaS3 DNA
