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Title: Molecular cloning, characterization and expression of a cDNA coding opper/zinc-superoxide dismutase from black porgy
Authors: Chi-Tsai Lin
Tung Liang Lee
Kow-Jen Duan
Chuian-Fu Ken.
Contributors: 國立臺灣海洋大學:生命科學系
Keywords: expression
Acanthopagrus schlegeli
Escherichia coli
Date: 2000
Issue Date: 2018-03-28T01:16:14Z
Publisher: Journal of Agriculture and Food Chemistry
Abstract: Abstract:A full-length complementary DNA (cDNA) clone encoding a putative copper/zinc superoxide dismutase (Cu/Zn-SOD) was amplified by a Polymerase Chain Reaction (PCR) based technique from cDNA synthesized from black porgy, Acanthopagrus schlegeli, mRNA. Nucleotide sequence analysis of this cDNA clone revealed that it comprised a complete open reading frame coding for 154 amino acid residues. The deduced amino acid sequence showed slightly higher identity (72.8−78.1%) with shark and swordfish Cu/Zn-SOD than with Cu/Zn-SOD from mammalian (68.1−70.7%) and plant (55.5−56.5%) sources. The residues required for coordinating copper and zinc are conserved as they are among all reported Cu/Zn-SOD sequences. The deduced amino acid sequence lacks mitochondria targeting sequence, which suggests that the black porgy cDNA clone encodes a cytosolic Cu/Zn-SOD. The coding region of Cu/Zn-SOD from black porgy was introduced into an expression vector, pET-20b(+), and transformed into Escherichia coli AD494(DE3)pLysS. A predominant achromatic zone was detected by activity staining of native PAGE. This indicates that the Cu/Zn-SOD cDNA clone can express active Cu/Zn-SOD enzyme in E. coli.
Relation: 48(9)
Appears in Collections:[Department of Life Science] Periodical Articles

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