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Volume 5, Issue 2 (Suppl)
Transcriptomics, an open access journal
ISSN: 2329-8936
Molecular Biology 2017
August 31-September 01, 2017
2
nd
International Conference on
August 31-September 01, 2017 Philadelphia, USA
Molecular Biology, Nucleic Acids &
Molecular Medicine
A CUGGU/UUGGU-specific MazF homologue from
Methanohalobium evestigatum
Yojiro Ishida
1
, Keiko Inouye
1
, Ouyang Ming
2
and
Masayori Inouye
1
1
Center for Advanced Biotechnology and Medicine, USA
2
University of Massachusetts, USA
MazF is a sequence-specific endoribonuclease or mRNA interferases, which cleaves RNA at a specific sequence. Since the
expression of a specific gene or a group of specific genes can be regulated by MazF, expanding the repertoire of recognition
sequences by MazF mRNA interferases is highly desirable for biotechnological and medical applications. Here, we identified
a gene for a MazF homologue (MazFme) from
Methanohalobium evestigatum
, an extremely halophilic archaeon. In order to
suppress the toxicity of MazFme to the E. coli cells, the C-terminal half of the cognate antitoxin MazEme was fused to the
N-terminal ends of MazFme. After purification of the MazEme-MazFme fusion protein, MazFme was released from the fusion
protein by factor Xa treatment. The free MazFme RNA cleavage specificity was determined by primer extension and synthetic
ribonucleotides, revealing that MazFme is a CUGGU/UUGGU-specific endoribonuclease.
Biography
Yojiro Ishida has recently obtained his PhD from Hiroshima University. His mentors are Professor Tadashi Shimamoto, Hiroshima University and Professor Masayori
Inouye, Rutgers University. He has developed a new expression system to incorporate a toxic amino acid analogue into a protein to alter the function by using
the single-protein production (SPP) system. Furthermore, he developed a residue and stereo specific labeling system for NMR structural studies. Currently, his
research focus are: Discovery of new MazF homologues, application of MazF for specific gene regulations, characterization of new Toxin-Antitoxin (TA) systems
from
Staphylococcus aureus
, and incorporating
19
F probe into methyl groups of a protein to characterize large molecular weight proteins and membrane proteins
using the SPP system in
E. coli
.
ishida@cabm.rutgers.eduYojiro Ishida et al., Transcriptomics 2017, 5:2 (Suppl)
DOI: 10.4172/2329-8936-C1-013