MacHattie L A, Shapiro J A
Proc Natl Acad Sci U S A. 1978 Mar;75(3):1490-4. doi: 10.1073/pnas.75.3.1490.
Phage lambdacam112, which contains the chloramphenicol resistance transposon Tn9 and has a deletion of attP and the int gene, will lysogenize Escherichia coli K-12. Prophage integration occurs at different chromosomal sites, including lacY and malB, but not at attB. All lambdacam112 prophages are excised from the chromosome after induction but with various efficiencies for different locations. Heteroduplex analysis of lambdaplacZ transducing phages isolated from a lacY::lambdacam112 prophage reveals an insertion sequence 1 (IS1) element at the joint of viral and chromosomal DNA. Two lines of evidence indicate that lambdacam112 encodes an excision activity that recognizes the IS1 element: (i) prophage derepression increases the frequency of excision from lacY to yield lac+ revertants, and (ii) lambdacam112 infection increases reversion of a galT::IS1 mutation about 50-fold. Our results indicate that the IS1 termini of TN9 can replace attP as a site for lambda insertion in the bacterial chromosome and that excision events are catalyzed by an IS1-encoded protein under lambda repressor and N gene control.
噬菌体λcam112含有氯霉素抗性转座子Tn9,且attP和int基因缺失,它能使大肠杆菌K-12溶源化。原噬菌体整合发生在不同的染色体位点,包括lacY和malB,但不在attB处。所有λcam112原噬菌体在诱导后都能从染色体上切除,但不同位置的切除效率各不相同。对从lacY::λcam112原噬菌体中分离出的λplacZ转导噬菌体进行异源双链分析,发现在病毒DNA与染色体DNA的连接处有一个插入序列1(IS1)元件。有两条证据表明λcam112编码一种能识别IS1元件的切除活性:(i)原噬菌体去阻遏增加了从lacY切除以产生lac+回复体的频率,(ii)λcam112感染使galT::IS1突变的回复频率增加约50倍。我们的结果表明,TN9的IS1末端可以取代attP作为λ插入细菌染色体的位点,并且切除事件是由在λ阻遏物和N基因控制下的IS1编码蛋白催化的。