Montgomery D L, Hall B D, Gillam S, Smith M
Cell. 1978 Jul;14(3):673-80. doi: 10.1016/0092-8674(78)90250-7.
The iso-1-cytochrome c gene of yeast has been identified and cloned using a synthetic oligodeoxynucleotide as a hybridization probe. The oligomer d[pT-T-A-G-C-A-G-A-A--C-C-G-G] is complementary to a region near the N terminal coding region of the yeast cyc 1 gene. Of several yeast Eco RI fragments which hybridize to this probe, one is changed in size by a G leads to T mutation which eliminates an Eco RI site within the cyc 1 gene. Both the wild-type and the RI- mutant forms were cloned in lambda gt vectors. Maxam-Gilbert sequencing for 91 nucleotides into the coding region for iso-1-cytochrome c yielded a DNA sequence in perfect correspondence with the known protein sequence.
酵母的同工-1-细胞色素c基因已通过使用合成寡脱氧核苷酸作为杂交探针进行了鉴定和克隆。寡聚物d[pT-T-A-G-C-A-G-A-A--C-C-G-G]与酵母cyc 1基因N端编码区附近的一个区域互补。与该探针杂交的几个酵母Eco RI片段中,有一个片段的大小因G突变为T而发生改变,该突变消除了cyc 1基因内的一个Eco RI位点。野生型和RI-突变型均克隆于λgt载体中。对同工-1-细胞色素c编码区的91个核苷酸进行Maxam-Gilbert测序,得到的DNA序列与已知蛋白质序列完全一致。