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Characterization of putative mutagenesis cassettes in Sinorhizobium meliloti (927.1)
Author(s) -
Genardi Samantha,
DeLateur Nicholas,
Kramer Caitlin,
Leifer Becky,
Brewer Tess,
Jones Kathryn,
Beuning Penny
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.927.1
Subject(s) - biology , mutagenesis , sinorhizobium meliloti , plasmid , dna polymerase , genetics , dna , gene , polymerase , dna replication , dna polymerase ii , transposon mutagenesis , microbiology and biotechnology , mutation , polymerase chain reaction , mutant , transposable element , reverse transcriptase
The goal of this research is to biochemically characterize chromosomal and plasmid‐encoded members of Y‐family DNA polymerases and putative mutagenesis cassettes from the nitrogen‐fixing plant symbiotic bacterium Sinorhizobium meliloti. Y‐family translesion polymerases are recruited to sites of DNA damage to replicate damaged DNA that may cause disruption to the replication fork. ImuC is a C family DNA polymerase with a putative function in mutagenic DNA replication; while ImuA and ImuB appear to be important for ImuC function, their roles have yet to be elucidated. S. meliloti harbors the Y family DNA polymerase dinB and the imuABC mutagenesis cassette encoded on both the chromosome and one of the symbiotic plasmids. All of the genes of the putative mutagenesis cassettes have been subcloned into two different plasmid vectors for in vivo and in vitro experiments. We are currently optimizing conditions for protein expression in E. coli . We are also carrying out assays to determine the ability of these gene products to confer on E. coli survival to a variety of DNA damaging agents and damage‐induced mutagenesis. Supported by NSF and ACS