Origin of the EcoRI Endonuclease G Fragment in the SP02 Bacteriophage Mutant Clh2 and Determination of the Temporal Order and Genomic Location of SP02 Bacteriophage Transcripts
Abstract
In developing Bacillus subtilis as a cloning host, phage for the
host should be isolated and characterized. A promising phage that can
be utilized as a cloning vector is bacteriophage SP02. SP02 has
deletion mutants and the host-range for the phage has been extended to
include Bacillus amyloliquefaciens and Bacillus natto. SP02 has been
shown to have several unique endonuclease sites and, more importantly,
there is a unique BglII endonuclease site in a genetically silent
region of the genome. Deletion mutants for bacteriophage SP02 have
been isolated. A host-range mutant of SP02, clh2 has an EcoRI digestion
fragment that is foreign to the SP02 DNA. The first goal of this
study was to determine the host with which SP02 DNA recombined to form
the clh2 mutant. Initial findings from this study show some homology
of biotinylated clh2 DNA to Bacillus amyloliquefaciens HSR chromosomal
DNA. A second goal of this study was to determine the temporal order
and genomic locations of SP02 transcripts. By hibridizing RNA
transcripts, isolated temporally, to digested SP02 DNA, results from
this study confirm a silent region in the SP02 genome, show temporal
ordering of the SP02 transcripts, and show the genomic location of the
SP02 transcripts. If you are not a current K College student, faculty, or staff member, email dspace@kzoo.edu to request access to this SIP.