Contreras R, Küpper H, Landy A, Khorana H G
J Biol Chem. 1976 Sep 10;251(17):5124-40.
The template-dependent primer elongation method for determining DNA sequences of specific regions (e.g. Loewen, P., Sekiaya, T., and Khorana, H. G. (1974) J. Biol. Chem. 249, 217-226) has been applied to the determination of the sequences of the promoters and of the regions beyond the C-C-A ends of the tyrosine tRNA genes in Escherichia coli. The following results have been obtained. (a) The promoter of the tRNA1Tyr (su+) gene in the bacteriophage phi80psu+III (the singlet strain) and phi80psu+-III (the doublet strain) and, significantly, the promoter of the tRNA2Tyr gene in the bacteriophage lambdah80dglyTsu+36 all have the following identical sequence in the first 59 nucleotides: (see article) Transcription begins at the underlined terminal nucleotide and proceeds to the right. (b) tRNA1Tyr (su+) as present in the singlet strain phi80psu+III and the tRNA1Tyr (su-), the second gene in the doublet strain phi80psu+-III, have the following identical sequence beyond the C-C-A sequences: (5') TCACTTCAAAAGTCCTGAACT (3') (c) tRNA1Tyr (su+), the first gene in the doublet strain phi80psu+-III, has the sequence (5') TAATTCACCACAGGG (CA) (3'), and tRNA2Tyr in lambdah80dglyTsu+36 has the sequence (5') ATTTCGGCCACGCGA (TGCGG) (3') beyond the C-C-A nucleotides.
用于确定特定区域DNA序列的模板依赖性引物延伸法(例如,Loewen, P., Sekiaya, T., 和Khorana, H. G. (1974) J. Biol. Chem. 249, 217 - 226)已应用于大肠杆菌中酪氨酸tRNA基因启动子序列以及C - C - A末端以外区域序列的测定。得到了以下结果。(a) 噬菌体phi80psu + III(单倍体菌株)和phi80psu + - III(双倍体菌株)中tRNA1Tyr(su +)基因的启动子,以及显著地,噬菌体lambdah80dglyTsu + 36中tRNA2Tyr基因的启动子,在前59个核苷酸中都具有以下相同序列:(见文章)转录从带下划线的末端核苷酸开始并向右进行。(b) 单倍体菌株phi80psu + III中存在的tRNA1Tyr(su +)以及双倍体菌株phi80psu + - III中的第二个基因tRNA1Tyr(su -),在C - C - A序列之后具有以下相同序列:(5') TCACTTCAAAAGTCCTGAACT (3') (c) 双倍体菌株phi80psu + - III中的第一个基因tRNA1Tyr(su +)具有序列(5') TAATTCACCACAGGG (CA) (3'),并且噬菌体lambdah80dglyTsu + 36中的tRNA2Tyr在C - C - A核苷酸之后具有序列(5') ATTTCGGCCACGCGA (TGCGG) (3') 。