摘要
用盐酸胍法和寡聚(dT)-纤维素亲和层析法从人胚肾细胞中提取了poly(A)-RNA,该poly(A)-RNA经麦胚无细胞体系测定其体外翻译活性,~3H-亮氨酸掺入量为空白对照的11.5倍。以此poly(A)-RNA为模板,合成了人胚肾细胞总cDNA,并用多聚物加尾法,与pUC19质粒重组,转化到E, Coli JM107中,进行分子克隆,其转化效率约为2×10~4克隆子/μg cDNA。
The poly(A)-RNA was extracted from human fetal kidneys by the guanidine-HCl method and purified by oligo(dT)-cellulose affinity chromatography. The in vitro translation activity of the poly(A)-RNA was asseyed in wheat germ system. The incorporation of 3H-leucine was 11.5 fold higher than control. The total cDNAs of human fetal kidneys were synthesized from the poly(A)-RNA. The cDNAs were inserted into pUC19 by homopolemeric tailing. The recombinant plasmids were transformed into E. coli JM107 with an efficiency about 2 × 104 colonies/μg cDNA.
出处
《北京大学学报（自然科学版）》
CAS
CSCD
北大核心
1989年第4期470-475,共6页
Acta Scientiarum Naturalium Universitatis Pekinensis
