, Volume 15, Issue 5, pp 637-640
Date: 05 Oct 2010

Construction of eukaryotic recombinant vector of renalase and its expression as a eukaryotic protein

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Abstract

Renalase is a secreted amine oxidase that metabolizes catecholamines. It has been proposed to modulate blood pressure and heart rate and its downregulation might result in hypertension. Despite its potential relevance for human health, the biochemical characterization of renalase is still scarce. The aim of this study is to synthesize the human renalase eukaryotic protein by genetic engineering. The human renalase gene was amplified by polymerase chian reaction (PCR). After digestion by BamH I and Xho I enzymes, the DNA fragments were cloned into the transfer vector, pFastBacHTb-Fc, to generate the pFastBacHTb-renalase expression vector. The ligation products were transformed into E. coli DH10Bac to obtain recombinant transposon rBacmid-renalase. The recombinant transposon was further transferred into insect high-V cells, and the recombinant human renalase eukaryotic protein was expressed successfully.

Foundation item: the Natural Scientific Research Project of Shanghai (No. 05ZR14086) and the Scientific Research Project of Shanghai Municipal Health Bureau (No. 2008Y034)