期刊
JOURNAL OF BIOCHEMISTRY
卷 147, 期 4, 页码 511-522出版社
OXFORD UNIV PRESS
DOI: 10.1093/jb/mvp197
关键词
catalase-peroxidise; furA-katG locus; Mn(II) binding; recombinant protein expression; transcription analysis
资金
- Korea Science and Engineering Foundation (KOSEF), Korea government (MEST) [R01-2008-000-12139-0]
- National Research Foundation of Korea [R01-2008-000-12139-0] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
The gene encoding a catalase-peroxidase (KatG) was cloned from chromosomal DNA of a fast-growing Mycobacterium sp. strain JC1 DSM 3803. The nucleotide sequence of a 5.7 kb EcoRI fragment containing the katG and its flanking regions was determined. The fragment (5,706 bps) contained two complete open reading frames (ORFs) encoding putative ferric uptake regulator A (FurA) and KatG proteins. The cloned gene, katG, had an ORF of 2241 nt, encoding a protein with calculated molecular mass of 81,748 Da. The furA was located in the upstream of the katG with the same transcriptional direction and there was a 38 bp gap space between them. The deduced KatG and FurA protein sequences showed significant homologies to KatG2 and Fur2 of Mycobacterium smegmatis and clustered with other mycobacterial KatG and Fur-like proteins in phylogenetic trees, respectively. The recombinant KatG overproduced in Escherichia coli was nearly indistinguishable from the native JC1 catalase-peroxidase in enzymatic properties and also possessed the resistance to organic solvents, indicating that the cloned katG truly encodes the Mycobacterium sp. JC1 catalase-peroxidase. Difference spectroscopy revealed Mn(II) binding near the haem of the KatG. Transcript analysis of the furA-katG using RT-PCR suggests that the katG is independently transcribed from the furA.
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