Isolation and characterization of ornithine decarboxylase gene from flounder (Paralichthys olivaceus).
Ornithine decarboxylase (ODC) is a homodimeric enzyme dependent on pyridoxal 5'-phosphate. We identified a complementary DNA clone corresponding to ODC from the brain of adult flounder (Paralichthys olivaceus). The flounder ODC cDNA consisted of 2939 bp encoding 272 amino acid residues. The flounder ODC showed 80.3% sequence identity to zebrafish and 70.8% to rat at the amino acid level. Comparison of the structure and nucleotide sequence of the ODC genes revealed that the gene is highly conserved in the flounder, zebrafish, and rat. The presence of ODC mRNA species in brain, kidney, liver, and embryo was confirmed using the reverse transcriptase polymerase chain reaction. The recombinant protein of flounder ODC containing a short histidine tag at the carboxyl terminus was overexpressed in Escherichia coli BL21 (DE3) codon plus using an inducible T7 expression system, and was purified by Ni-NTA affinity chromatography.
Lee
Jae Hyung
JH
Department of Microbiology, Pukyong National University, Busan 608-737, Korea. ytkim@pknu.ac.kr
Son
Mi Young
MY
Yoon
Moon-Young
MY
Choi
Jung-Do
JD
Kim
Young Tae
YT
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000201:enzymology; D002846:Chromatography, Affinity; D016000:Cluster Analysis; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004625:Embryo, Nonmammalian / Q000201:enzymology; D005432:Flounder / Q000235:genetics*; D015870:Gene Expression; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D009955:Ornithine Decarboxylase / Q000235:genetics*; D010802:Phylogeny; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D009955:Ornithine Decarboxylase