Agrigenetics Advanced Research Division, 5649. E. Buckeye Road, 53716, Madison, WI, USA.
Plant Mol Biol. 1983 Nov;2(6):335-50. doi: 10.1007/BF01578595.
The complete nucleotide sequence of the transferred region (T-DNA) of an octopine tumor inducing (Ti) plasmid fromAgrobacterium tumefaciens (pTi15955) has been determined. A total of 24 595 nucleotides extending approximately 900 bases to either side of the outermost, T-DNA boundaries was sequenced. Computer analysis of the sequenced portion of the Ti plasmid revealed that recognition sites for 72 restriction endonucleases are present in the DNA sequence at least once; no site forEcoK exists in this DNA sequence. Two imperfect 24 base repeats border the T-DNA sequence; the left starts at position 909 and the right ends at position 23 782, giving the T-DNA region a total length, of 22 874 nucleotides. Another two similar 24 base repeats lie within T-DNA and divide it, into three distinct domains: T-left (TL-DNA) 13 175 bp of apparently eukaryotic origin; T-center (TC-DNA) 1816 bp of prokaryotic origin; and T-right (TR-DNA) 7 883 bp of eukaryotic origin. The T-DNA contains nine reported transcripts, however, 26 open reading frames longer than 300 bases that start with an ATG initiation codon were found. Fourteen open reading frames are bounded by putative eukaryotic promoters, ribosome binding sites, and poly(A) addition sites and occur only in TL-and TR-DNAs. No open reading frames showing eukaryotic promoter sequences are located within the TC-DNA.
已测定根癌农杆菌(Agrobacterium tumefaciens)(pTi15955)的章鱼碱肿瘤诱导(Ti)质粒的转移区(T-DNA)的完整核苷酸序列。总共测序了大约 900 个碱基,延伸到 T-DNA 边界最外侧的 24595 个核苷酸。Ti 质粒测序部分的计算机分析显示,该 DNA 序列中至少存在 72 种限制内切酶的识别位点;该 DNA 序列中不存在 EcoK 位点。T-DNA 序列两侧有两个不完整的 24 碱基重复;左侧从第 909 位开始,右侧在第 23782 位结束,使 T-DNA 区域的总长度为 22874 个核苷酸。另外两个类似的 24 碱基重复位于 T-DNA 内,并将其分为三个不同的区域:T-左侧(TL-DNA)13175 个碱基,显然来自真核生物;T-中心(TC-DNA)1816 个碱基,来自原核生物;T-右侧(TR-DNA)7883 个碱基,来自真核生物。T-DNA 包含九个报告的转录本,但发现了 26 个长于 300 个碱基的开放阅读框,其起始密码子为 ATG。有 14 个开放阅读框被假定的真核启动子、核糖体结合位点和 poly(A)添加位点所限制,仅出现在 TL 和 TR-DNAs 中。在 TC-DNA 中没有发现具有真核启动子序列的开放阅读框。