Padmanabhan R, Jay E, Wu R
Proc Natl Acad Sci U S A. 1974 Jun;71(6):2510-4. doi: 10.1073/pnas.71.6.2510.
We have developed a general approach for determining the nucleotide sequence of a gene, with the aid of a deoxyribonucleotide primer of defined sequence. The selection of the primer sequence was based on a short segment of mRNA sequence of T4 phage lysozyme. A tetradecadeoxyribonucleotide primer was chemically synthesized and its sequence verified by sequence analysis. This primer was found to bind to the single-stranded region of the exonuclease III-treated T4 DNA, and specific nucleotides were incorporated to its 3' end. The result indicated that this primer was bound to the expected location on the T4 DNA. Therefore, long sequences of the T4 lysozyme gene can now be determined from this specific starting point.
我们借助一段已知序列的脱氧核糖核苷酸引物,开发出了一种确定基因核苷酸序列的通用方法。引物序列的选择基于T4噬菌体溶菌酶mRNA序列的一小段。化学合成了一个十四聚体脱氧核糖核苷酸引物,并通过序列分析验证了其序列。发现该引物可与经核酸外切酶III处理的T4 DNA的单链区域结合,特定的核苷酸被掺入其3'末端。结果表明该引物与T4 DNA上预期的位置结合。因此,现在可以从这个特定的起始点确定T4溶菌酶基因的长序列。