A study on cloning and expression of a fragment of DFNB59 gene encoding human pejvakin in Escherichia coli

Nguyen Thi Kim Cuc, Kazutada Watanabe, Manabu Toyoshima, Yasushi Shimoda

Abstract


DFNB59 is a newly identified gene on chromosome 2q31.1q31.3. The mutations on DFNB59 involved human nonsyndromic deafness. DFNB59 encodes pejvakin, a 352-residue protein. Pejvakin is a paralog of DFNA5, a protein of unknown function of the gasdermin family also involved deafness [3]. In this research, pBluescript II SK was used as the template for amplification of the fragment f1DFNB59. This fragment was then cloned in pGEX-3X and transformed to E. coli Top 10 for storage. The recombinant plasmids pGEX-3X containing f1DFNB59 from transformations of E. coli Top 10 was transformed to E. coli BL21 for expression of the GST-fusion protein. This GST-fusion proteins was purified by affinity chromatography using immobilized Glutathione. The protein (>40 kDa) obtained after purification can be used as an antigen for production of monoclonal antibodies against pejvakin.

Keywords


DFNB59 gene, GST-fusion protein, nonsyndromic deafness, pejvakin proteins.

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DOI: https://doi.org/10.15625/0866-7160/v36n1se.4368

Published by Vietnam Academy of Science and Technology