Gillam S, Smith M
Gene. 1979 Dec;8(1):81-97. doi: 10.1016/0378-1119(79)90009-x.
A synthetic oligodeoxyribonucleotide mismatched at a single nucleotide to a specific complementary site on wild-type circular phi X174 DNA can be used to produce a defined point mutation after in vitro incorporation into closed circular duplex DNA by elongation with DNA polymerase and ligation followed by transfection of Escherichia coli (Hutchison et al., 1978; Gillam et al., 1979). The present study is an investigation of the optimum conditions required for the oligodeoxyribonucleotide-primed reaction for production of transition and transversion mutations in phi X174 DNA, using the large (Klenow) fragment of E. coli DNA polymerase I. Under optimum conditions up to 39% of the progeny of transfection are the desired mutant and significant mutation is observed using a heptadeoxyribonucleotide.
一种在单个核苷酸处与野生型环状φX174 DNA上的特定互补位点错配的合成寡脱氧核糖核苷酸，在通过DNA聚合酶延伸并连接后体外掺入闭环双链DNA中，随后转染大肠杆菌后，可用于产生特定的点突变（哈钦森等人，1978年；吉勒姆等人，1979年）。本研究使用大肠杆菌DNA聚合酶I的大片段（克列诺片段），对在φX174 DNA中产生转换和颠换突变的寡脱氧核糖核苷酸引发反应所需的最佳条件进行了研究。在最佳条件下，高达39%的转染子代是所需的突变体，并且使用十七脱氧核糖核苷酸可观察到显著的突变。
