Molecular cloning, characterization, and expression of a cDNA coding copper/zinc superoxide dismutase from black porgy

Chi Tsai Lin, Tung Liang Lee, Kow Jen Duan, Chuian Fu Kent

Research output: Contribution to journalArticle

13 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)4444-4447
Number of pages4
JournalJournal of Agricultural and Food Chemistry
Volume48
Issue number9
DOIs
Publication statusPublished - 2000 Oct 13

Fingerprint

porgy
complementary DNA
Cloning
Molecular Cloning
Superoxide Dismutase
Zinc
Copper
molecular cloning
superoxide dismutase
Complementary DNA
copper
zinc
Clone Cells
Amino Acids
Escherichia coli
Amino Acid Sequence
clones
Native Polyacrylamide Gel Electrophoresis
Sharks
Mitochondria

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Agricultural and Biological Sciences(all)

Cite this

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title = "Molecular cloning, characterization, and expression of a cDNA coding copper/zinc superoxide dismutase from black porgy",
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.",
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Molecular cloning, characterization, and expression of a cDNA coding copper/zinc superoxide dismutase from black porgy. / Lin, Chi Tsai; Lee, Tung Liang; Duan, Kow Jen; Kent, Chuian Fu.

In: Journal of Agricultural and Food Chemistry, Vol. 48, No. 9, 13.10.2000, p. 4444-4447.

Research output: Contribution to journalArticle

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