Identification and characterization of a novel 1-Cys peroxiredoxin from silkworm, Bombyx mori.
Peroxiredoxins (Prxs) are believed to play an important role in insects for protection against the toxicity of reactive oxygen species (ROS). A gene encoding a novel 1-Cys Prx was firstly identified and characterized from an expressed sequence tag database (EST) in the lepidopteran insect, Bombyx mori. The 1-Cys Prx of B. mori (BmPrx) cDNA contained an open reading frame of 672 bp encoding a protein of 223 amino acid residues with calculated molecular mass of 25 kDa and included conserved cysteine residues signature motifs (PVCT) in the N-terminus amino acid. Sequence comparison showed that BmPrx shared 53 to 64% identity with other species 1-Cys proteins. RT-PCR revealed that the BmPrx transcripts were present in all tissues and developmental stages. The coding sequence was cloned and expressed as a 30-kDa protein in Escherichia coli. The purified enzyme acted as a catalyst in ferrithiocyanate system and protected supercoiled form of plasmid DNA from damage in metal-catalyzed oxidation (MCO) system in vitro. In addition, real-time PCR analysis indicated that significant rise of the transcripts level of BmPrx was induced by temperature stress including low and high temperature stimuli.
Wang
Qiang
Q
Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, China.
Chen
Keping
K
Yao
Qin
Q
Zhao
Yuan
Y
Li
Yijia
Y
Shen
Hongxing
H
Mu
Runhong
R
D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D030541:Databases, Genetic; D004926:Escherichia coli / Q000235:genetics; D020224:Expressed Sequence Tags / Q000737:chemistry / Q000378:metabolism; D015870:Gene Expression; D019476:Insect Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D054464:Peroxiredoxins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D019476:Insect Proteins; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D054464:Peroxiredoxins