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DNA SEQUENCE ANALYSIS OF BACILLUS PHAGE PHI29 RIGHT EARLY REGION AND LATE GENES 14, 15 AND 16 (LYSOZYME).

机译:杆菌噬菌体PHI29右早期区域和后基因14、15和16(溶菌酶)的DNA序列分析。

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摘要

The sequence of the rightmost 4,626 bp of the Bacillus phage φ29 genome is presented and analyzed. Nine large open reading frames (ORF's) have been found. Three of these ORF's are correlated with the late genes 14, 15 and 16. The remaining six ORF's are in the right early region. One of these early ORF's has been identified as gene 17 (g17), the only early gene to have been genetically mapped in this region. The remaining ORF's (16.5, 16.6, 16.7, 16.8 and 16.9) were previously unknown. The biological efficacies of some of these putative early ORF's were demonstrated using an in vitro E. coli transcription-translation system. The primary amino acid sequences, molecular weights, translational initiation sequences and genetic organization of these nine genes are presented and discussed. Gene product 15 (gp15) was found to have strong homology with Salmonella phage P22 gp19, a lysozyme. gp15 also has a lesser but possibly significant homology with T4 gene product e (gpe), also a lysozyme. Using a clone containing φ29 g15 it was shown that gp15 can complement T4 gene e (ge) mutant infections, leading to the conclusion that φ29 g15 encodes a lysozyme. Three transcriptional initiation sites (P(E)3, P(EC)3 and B2) were previously mapped in this region. The sequences of the putative P(EC)3 and B2 promoter sites are presented and shown to have homology with the Bacillus σ⁵⁵ concensus sequence. Sequences having homology to a minor Bacillus sigma factor recognition site, σ³², are also presented and discussed. The region between the last late gene (g16) and the last early gene (ORF-16.5) consists of only 30 bp. Analysis of potential secondary structures of transcripts across this region suggests that the same sequences may be involved in the termination of both late and early transcription.
机译:呈现并分析了芽孢杆菌噬菌体φ29基因组最右边4,626 bp的序列。找到了九个大型开放阅读框(ORF)。这些ORF中的三个与晚期基因14、15和16相关。其余六个ORF在正确的早期区域。这些早期ORF之一已被鉴定为基因17(g17),这是在该区域进行了遗传定位的唯一早期基因。其余的ORF(16.5、16.6、16.7、16.8和16.9)以前是未知的。使用体外大肠杆菌转录-翻译系统证明了其中一些早期ORF的生物功效。介绍并讨论了这9个基因的主要氨基酸序列,分子量,翻译起始序列和遗传组织。发现基因产物15(gp15)与溶菌酶沙门氏菌噬菌体P22gp19具有强同源性。 gp15与T4基因产物e(gpe)(也是一种溶菌酶)的同源性也较低,但可能很重要。使用含有φ29g15的克隆显示gp15可以补充T4基因e(ge)突变感染,从而得出结论,φ29g15编码溶菌酶。三个转录起始位点(P(E)3,P(EC)3和B2)先前已在此区域定位。提出了假定的P(EC)3和B2启动子位点的序列,并显示与芽孢杆菌σ共有序列具有同源性。还介绍并讨论了与次要芽孢杆菌西格玛因子识别位点σ³²具有同源性的序列。最后一个晚期基因(g16)和最后一个早期基因(ORF-16.5)之间的区域只有30 bp。跨该区域的转录物的潜在二级结构的分析表明,相同的序列可能参与晚期和早期转录的终止。

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    GARVEY KEVIN JAMES.;

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  • 年度 1986
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