Isolation of lambda transducing phage with the bio genes inserted between lambda genes P and Q.
Isolation of lambda transducing phage with the bio genes inserted between lambda genes P and Q.
复制标题
分离 lambda 转导噬菌体，其中 bio 基因插入 lambda 基因 P 和 Q 之间。
DOI：
10.1093/genetics/95.1.1
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发表时间：
1980
期刊：
影响因子：
3.3
通讯作者：
Stamm,SC
中科院分区：
文献类型：
--
作者：
Charon,NW;Campbell,AM;Stamm,SC
在λ基因P和q之间插入生物基因，构建了形成斑块的生物素转导噬菌体。这些噬菌体的最终目的是将λQ基因融合到生物操纵子上。利用λ基因βNcI857OPQSRA左侧邻近的生物基因构建了一个有缺陷的温度敏感溶解菌原。对耐热幸存者进行筛选，以确定终点在生物操纵子和λP右侧和λA左侧的缺失。在大约1600名耐热幸存者中，有5人具有这些特性。其中两人的基因序列为bioAB....λQSRA。用λcI857b221溶原并热诱导获得所需的转导噬菌体。我们对这些噬菌体进行了表征，并对其中一种进行了详细的研究。三分之二分离的斑块形成转导噬菌体携带整个bioB基因和部分bioA基因，三分之一携带整个bioA和bioB基因。分离的噬菌体在繁殖过程中失去了生物基因，表明它们含有部分噬菌体基因的复制。结果表明，在其中一个噬菌体λbioq1b221中，该重复不涉及整个λQ基因。该噬菌体重组体λNam7am53c17bioq1b221在生物素抑制条件下无法形成斑块。我们得出结论，如果λQ基因与该噬菌体的生物操纵子融合，则没有足够的λQ基因产物允许噬菌体繁殖。
Plaque-forming, biotin-transducing phages were constructed with the bio genes inserted between lambda genes P and Q. These phages were isolated for the eventual aim of fusing the λQ gene to the bio operon. The following steps were used to construct these phages: A defective temperature-sensitive lysogen was constructed with the bio genes adjacent to and to the left of lambda genes βNcI857OPQSRA. Heat-resistant survivors were screened for deletions with endpoints in the bio operon and to the right of λP and to the left of λA. Five of approximately 1,600 heat-resistant survivors had these properties. Two had the gene order bioAB.... λQSRA. When these two strains were lysogenized with λcI857b221 and heat induced, the desired transducing phages were obtained. We characterized these phages and studied one in detail. Two-thirds of the plaque-forming transducing phages isolated carried the entire bioB gene and only part of the bioA gene, and one-third carried the entire bioA and bioB genes. The phages isolated lost the bio genes upon propagation, indicating that they contain a partial duplication of phage genes. The duplication was shown not to involve the entire λQ gene in one of these phages, λbioq1b221. A recombinant of this phage, λNam7am53c17bioq1b221, failed to form plaques under biotin-derepression conditions. We conclude that if the λQ gene was fused to the bio operon in this phage, not enough λQ gene product was made to allow phage propagation.