Correlated genetic and EcoRI cleavage map of Bacillus subtilis bacteriophage phi105 DNA

Abstract

The seven previously identified EcoRI cleavage fragments of 4105 DNA were ordered with respect to their sites of origin on the phage genome by marker rescue. One fragment, H, did not carry any determinants essential for replication. This fragment was totally missing in a deletion mutant which exhibited a lysogenization-defective phenotype. There is a nonessential region on the 4105 genome which begins in fragment B, spans fragment H, and ends in fragment F. The size of the nonessential region, as estimated by alterations observed in the fragmentation patterns of deletion mutant DNAs, is approximately 2.7 x 106 daltons. Two new EcoRI cleavage fragments with molecular weights of approximately 0.2 x 106 were detected by autoradiography of 32P-labeled DNA. These small fragments were not located on the cleavage map. MATERIALS AND METHODSPhage and bacterial strains. The wild-type and conditionally lethal mutant strains of 0105 were from 395 on August 1, 2020 by guest

Cloning restriction fragments that promote expression of a gene in Bacillus subtilis

“…EcoRI* or EcoRI activity (15) was used to digest pPL10, pUB110, and DNA from B. Iicheniformis 9945A and phage SP02 (10,13,17,19). Approximately 1 ug of EcoRI*or EcoRIdigested DNA was mixed with 1 ,ug of EcoRIdigested pPL603 in 100 P1.…”

Restriction endonuclease mapping of bacteriophage phi105 and closely related temperate Bacillus subtilis bacteriophages rho10 and rho14

“…Large capital letters identify EcoRI fragments for each phage DNA, as shown in Table 3. Small capital letters associated with the q0105 fragments indicate cistrons which occur on those fragments (Scher et al,[19]). The region indicated by lines lettered DI:1c, DI:4c, DI:2c, DI:lt, DI:29t, and DII:6c demonstrate deletion mutants in the 4105 genome(4).…”

Transfection ofBacillus subtiliswith phage φ105 DNA: Effect of φ105EcoRI restriction endonuclease fragments

“…In marker rescue with 0105 susL10 as superinfecting phage 0.1 /~g fragment gave about 105 infectious centers. Gene L of 0105 is located on the EcoRI-C fragment which is part of the A fragment [5]. Rescue of the L gene from the A fragment is more efficient compared to mature DNA possibly due to internalization of the cohesive ends (data not shown).…”

“…Experimental support for this suggestion has been provided by the finding that when the cohesive ends are protected by additional DNA such as in prophage DNA extracted from lysogenic bacteria, in concatemers formed during phage DNA replication, or in oligomers formed by in vitro treatment of mature DNA with DNA ligase a single molecule is infectious and the efficiency of transfection is increased manyfold [4]. Restriction endonuclease RI cleaves mature q~105 DNA into eight fragments [5,6]. The cohesive ends are located in fragments C (5.8…”