噬菌体T4抗限制内切核酸酶基因的物理图谱绘制与克隆
Physical mapping and cloning of bacteriophage T4 anti-restriction endonuclease gene.
作者信息
Dharmalingam K, Revel H R, Goldberg E B
出版信息
J Bacteriol. 1982 Feb;149(2):694-9. doi: 10.1128/jb.149.2.694-699.1982.
Abstract
We have proposed that the ability of T4 to produce non-glucosylated progeny after a single cycle of growth on a galU rglA rglB+ mutant of Escherichia coli is due to the initiation of the rglB+ function by a phage-coded, anti-restriction endonuclease protein. Based on this hypothesis, we screened T4 deletion mutants for failure to give a burst in this host. The absence of an arn gene in phage mutants lacking the 55.5- to 58.4-kilobase region is verified by their inability to protect secondary infecting non-glucosylated phage from rglB-controlled cleavage. A functional arn gene was cloned on plasmid pBR325, and the 0.8-kilobase insert DNA was shown to be homologous to the DNA missing in the arn deletion phage.
摘要
我们提出，T4在大肠杆菌galU rglA rglB +突变体上经过一个生长周期后产生非糖基化子代的能力，是由于噬菌体编码的抗限制内切酶蛋白启动了rglB +功能。基于这一假设，我们筛选了T4缺失突变体，以确定其在该宿主中无法产生爆发。通过缺乏55.5至58.4千碱基区域的噬菌体突变体中arn基因的缺失，验证了它们无法保护二次感染的非糖基化噬菌体免受rglB控制的切割。一个功能性arn基因被克隆到质粒pBR325上，并且0.8千碱基的插入DNA被证明与arn缺失噬菌体中缺失的DNA同源。
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