Mutant of Lambda Bacteriophage Producing a Thermolabile Endolysin

Abstract

, N.Y.) AND ALICE DEL CAMPILLO-CAMPBELL. Mutant of lambda bacteriophage producing a thermolabile endolysin. J. Bacteriol. 85:1202-1207. 1963.-Endolysin from lambda bacteriophage and a temperature-sensitive mutant thereof was partially purified. The mutant enzyme was distinguishable from the wild type by its greater rate of inactivation by high tem

High-Throughput Characterization of Cascade type I-E CRISPR Guide Efficacy Reveals Unexpected PAM Diversity and Target Sequence Preferences

“…(B) Graphical depiction of the crRNA and DNA complex. The colors indicate the PAM (red), seed (blue), and distal (green) regions of the target-crRNA studies, we chose a temperature-sensitive mutant in the phage R gene ( (Campbell and Del Campillo-Campbell 1963) the l-Rts phage was a generous gift from Allan Campbell). A bacterial lawn was made with 100 ml of the overnight culture mixed with 5 ml of 0.7% top agar and spread on an LB plate.…”

“…All high-throughput retention assay data are deposited at the National Center for Biotechnology Information (NCBI) Archive (Study Accession PRJNA388730 (SRP108442)). Provisional "working model" sequence assemblies for plasmid pPD207.846 (File S1) and for the assembled l-Rts genome (File S2) (Campbell and Del Campillo-Campbell 1963) are provided with this manuscript as Supplemental Material.…”

“…We designed the in vivo high-throughput spacer screen with the intention of assaying a large population of crRNAs with varying PAMs and sequence characteristics. To develop an assay that would meet these factors, we designed a crRNA library that targeted the l-Rts phage genome (Campbell and Del Campillo-Campbell 1963). Sequences tiling the l genome with a window of 32 nts were generated; 12,472 sequences were selected and synthesized via massively parallel solid phase oligo synthesis ( Figure 1A).…”

“…In order for the spacer library to have the potential of self-removal by self-targeting, the cloned spacer library was transformed into a previously characterized bacterial strain that is recombination deficient with one copy of the Cascade CRISPR-Cas system (cas3ABCDE) on an arabinose promoter, also known as the ACT-01 strain (Caliando and Voigt 2015). An integrated l bacteriophage (l-Rts) (Campbell and Del Campillo-Campbell 1963) ( Figure 1A) has been added to the genome through lysogeny to allow targeting by l-derived spacer sequences. To obtain precise information on the target, we resequenced the genome of the l-Rts used to produce the target lysogen (File S2).…”

Stimulation of genetic recombination among lambda bacteriophages by ultraviolet radiation: The mechanism and the relevance of DNA repair processes

“…The B cistron product is necessary for the conversion of freshly synthesized phage DNA to an infectious form, and the G cistron function is required for the production of functional phage tails (232). The R cistron codes for the phage lysozyme (34). ~susQ73 is defective in a cistron that supplies a general inducer of late phage functions, 'vhile ~sus029 and ~susP3 are mutant in functions essential to vegetative DNA replication (29,48,121).…”