cDNA cloning and overexpression of an isoperoxidase gene from Korean-radish, Raphamus sativus L.

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Title
cDNA cloning and overexpression of an isoperoxidase gene from Korean-radish, Raphamus sativus L.
Author(s)
Jong Hoon Park; Soung Soo Kim
Bibliographic Citation
BMB Reports, vol. 29, no. 2, pp. 137-141
Publication Year
1996
Abstract
A partial cDNA encoding a Korean radish isoperoxidase was obtained from a cDNA library prepared from 9 day old radish root. In order to obtain Korean radish isoperoxidase cDNA, 5′ RACE (rapid amplification cDNA end) PCR was performed and a cDNA (prxK1) encoding a complete structural protein was obtained by RT (reverse transcription)-PCR. Sequence analysis revealed that the length of the cDNA was 945 base pairs, and that of the mRNA transcript was ca. 1.6 kb. The deduced amino acid of the protein were composed of 315 amino acid residues and the protein was 92% homologous to turnip peroxidase, and 46% to 50% homologous to other known peroxidases. The 945 bp cDNA encoding Korean radish isoperoxidase was overexpressed in Escherichia coli up to approximately 9% of total cellular protein. The recombinant fusion protein exhibited 43 kDa on SDS-PAGE analysis and the activity level of the recombinant nonglycosylated protein was two fold higher in IPTG induced cell extracts than that of uninduced ones.
Keyword
cDNA cloning5′ RACE (rapid amplification cDNA end) and RT (reverse transcription)-PCRoverexpression
ISSN
1225-8687
Publisher
South Korea
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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