Correlation between genetic map and map of cleavage sites for sequence-specific endonucleases SalI, KpnI, BglI, and BamHI in bacteriophage T4 cytosine-containing DNA

A complete map of the cleavage sites of restriction endonucleases BamHI, BglII, KpnI, PvuI, SalI, and XbaI was determined for the cytosine-containing DNA of a bacteriophage T4 alc mutant. The 56 sequence-specific sites were assigned map coordinates based on a least-squares analysis of measured fragment lengths. Altogether, the lengths of 118 fragments from single and double enzyme digestions were measured by electrophoresis of the fragments in agarose gels. DNA fragments of known sequence or DNA fragments calibrated with fragments of known sequence were used as standards. The greatest deviation between an experimentally measured fragment length and its computed map coordinates was 3.0%; the average deviation was 0.8%. The total length of the wild-type T4 genome was calculated to be 166,200 base pairs.

mentioning

confidence: 99%

Map of restriction sites on bacteriophage T4 cytosine-containing DNA for endonucleases bamHI, BglII, KpnI, PvuI, SalI, and XbaI

Marsh¹,

Hepburn²

1981

“…Since the isolation of bacteriophage T4 mutants which allow the growth and packaging of DNA containing cytosine instead of the glucosylated hydroxymethylcytosine residues normally present (15), T4 DNA has been amenable to analysis with restriction endonucleases. Circular cleavage maps of the 166-kilobase pair T4 genome have been published for restriction enzymes BamHI, BglII, KpnI, SalI, SmaI, and XhoI (3,6,14), and maps for BglI, EcoRI, HindIII, and PstI have been completed recently (2,12). In the accompanying report, the sites of cleavage by PvuI and XbaI were added to a computer-fitted, unified map of the BamHI, Bglll, KpnI, and SailI cleavage sites (9).…”

mentioning

confidence: 99%

“…O'Farrell et al (12) have independently aligned a T4 restriction map with the genetic map, primarily by filter hybridization of restriction fragments to cloned segments of T4 DNA known to carry specific genes. Also, Carlson (2) has determined a general alignment based on the cleavage patterns of cytosine-containing DNA from T4 alc mutants which carry various characterized deletions. Our results are in agreement.…”

mentioning

confidence: 99%

Alignment of a restriction map with the genetic map of bacteriophage T4

Yee¹,

Marsh²

1981

A restriction map of the bacteriophage T4 genome was aligned with the T4 genetic map. Included were the cleavage sites for BamHI, BglII, KpnI, PvuI, Sall, and XbaI. The alignment utilized the fact that the T4 genetic map had been oriented previously with respect to a T2/T4 heteroduplex map. DNA fragments from a BgII digestion of cytosine-containing DNA from a T4 dCTPase-denA denB(rIIH23B) alc mutant were hybridized with full-length chromosomal strands of bacteriophage T2, and the heteroduplexes were examined by electron microscopy. From their lengths and patterns of substitution and deletion loops, the heteroduplexes formed with 6 of the 13 BglIl fragments could be unambiguously identified and positioned on the T2/T4 heteroduplex map. The ends of the T4 DNA strands in the heteroduplexes directly identified the location of 10 BglII cleavage sites. The remaining three BglII cleavage sites could be assigned to the T2/T4 heteroduplex map based on their relative locations on the restriction map. It was also possible to identify the source of the DNA strands (i.e., T2 or T4) in four previously unassigned deletion loops on the T2/T4 heteroduplex. Among the BglII fragments identified in heteroduplexes was the fragment containing the rIIH23B deletion; this deletion was used as the primary point of reference for alignment of the T4 restriction map with the T2/T4 heteroduplex map and, hence, with the T4 genetic map. Snyder (15). Originally, this T4 alc mutant was thought to carry the 4.2-kb rIIH23 deletion, but instead it has been found to carry a longer rII deletion, defined genetically as extending from or near the end 115 on July 6, 2020 by guest

“…The presence of denA or denB mutations in multiple mutants containing 56mutations was confirmed by complementation assays in liquid culture as follows: denA-, lack of ability to rescue phage production in mixed infection of E. coli B with denA-,55amC64X5 in the presence of 0.01 M hydroxyurea (19); denB-, high levels of DNA synthesis in mixed infection of E. coli B with denBD2a23 x 5, 56or with denBD2a2xS,56- (8,51). T4 Cyt-O is a pentuple mutant that makes viable phage containing fully cytosine-substituted T4 DNA when grown on E. coli B834 (9).…”

Section: Strains Of Bacteria and Bacteriophage E Coli B (14)mentioning

confidence: 99%

“…Hybridization. Nick translation (31), blotting (49), and hybridization were carried out as described previously (9).…”

Section: Strains Of Bacteria and Bacteriophage E Coli B (14)mentioning

confidence: 99%

In vivo cleavage of cytosine-containing bacteriophage T4 DNA to genetically distinct, discretely sized fragments

Carlson¹,

Wiberg²

1983

Self Cite

Source" and reference 42 amN55 x 5 dCMP hydroxymethylase-This laboratory 42 amN122 x 5 dCMP hydroxymethylase-This laboratory 46 amB14 x 5 46-47 nuclease-This laboratory 46 amN126 46-47 nuclease-This laboratory 56 amE51 x 5 dCTPase-; Cyt This laboratory 56 amE114 x 5 dCTPase-; Cyt This laboratory denA S112 x 5 endonuclease 11-This laboratory (19) denA SP4 x 5 endonuclease lI-This laboratory (19) denB D2a2 x 5 endonuclease IV-Vetter and Sadowski (51) denB D2a23 x 5 endonuclease IV-Vetter and Sadowski (51) denB saA&9 endonuclease IV-(12)