Rougeon F, Kourilsky P, Mach B
Nucleic Acids Res. 1975 Dec;2(12):2365-78. doi: 10.1093/nar/2.12.2365.
Double stranded DNA has been synthesized in vitro from rabbit globin messenger RNA and elongated with homopolymeric dG tails. An E. coli plasmid was cleaved by EcoRI. The cohesive ends were repaired and dC tails added, to permit reconstitution of the EcoRI sites upon annealing with the dG elongated globin DNA. Transformation of E. coli with the globin-plasmid DNA hybrid has yielded a clone which harbours a recombinant plasmid (pCR1-betaG1), as demonstrated by hybridization experiments with radioactive globin cDNA. The sequence carried by the recombinant plasmid corresponds to part of the gene sequence coding for the beta chain of rabbit globin. Circular DNA of the purified recombinant plasmid exhibits sensitivity to EcoRI.
已从兔珠蛋白信使核糖核酸体外合成双链DNA，并接上同聚物dG尾进行延伸。用EcoRI切割大肠杆菌质粒。修复黏性末端并添加dC尾，以便与dG延伸的珠蛋白DNA退火时恢复EcoRI位点。用珠蛋白-质粒DNA杂种转化大肠杆菌产生了一个克隆，该克隆含有重组质粒（pCR1-βG1），这通过与放射性珠蛋白cDNA的杂交实验得以证明。重组质粒携带的序列对应于编码兔珠蛋白β链的部分基因序列。纯化的重组质粒的环状DNA对EcoRI敏感。
