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Study of Methyletransferase RumB in E. coli
Author(s) -
Pritts Jordan,
Zilinskas Egidijus
Publication year - 2015
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.29.1_supplement.711.12
Subject(s) - rna , ribosomal rna , uracil , microbiology and biotechnology , protein subunit , ribosomal protein , methyltransferase , lysis , 23s ribosomal rna , biology , plasmid , chemistry , ribosome , biochemistry , gene , dna , methylation
In this experiment methyltransferases RumA and RumB were studied. These proteins are part of a group of enzymes that conduct post transcriptional modifications on RNA. RumB specifically methylates uracil 747 on the 23S subunit of ribosomal RNA. RumA methylates uracil 1939 on the 23S subunit of ribosomal RNA. Both RumA and RumB transfer a methyl group from a donor molecule (S‐Adenosylmethionine) to a receptor molecule (uracil on RNA). These two proteins share significant similarity in sequence as well as the incorporation of an Fe‐S cluster. Due to the similarity it is proposed that RumB should follow RumA characteristics. The RumA and RumB genes were successfully amplified using PCR, ligated into a plasmid, transformed, sequenced, and induced. The proteins were expressed using the pET‐45b expression vector. Cell lysis was not successful as the protein of interest remained in the cell pellet as opposed to the supernatant. Future research will include protein purification and analysis.