Improvement of the Exon Method for Rapid Synthesis of cDNA of the Rat Renalase Gene

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Published: Oct 12, 2023
DOI: https://doi.org/10.18097/BMCRM00201
Keywords:
gene expression; PCR; exon assembly; exon; rat renalase gene

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V.I. Fedchenko
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
A.A. Kaloshin
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia
A.E. Medvedev
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119121 Russia

Abstract

We have improved our previously developed method of exon cloning of cDNA of eukaryotic genes to obtain the rat renalase gene cDNA. In contrast to the previously used step-by-step pairwise assembly of exons, in this work the procedure of full-length cDNA preparation was shortened due to simultaneous assembly of four neighboring exons at once (exons 1-4 and exons 6-9 of the rat renalase gene). The two obtained sequences (exons 1-4 and 6-9) were combined into a full-length cDNA of the rat renalase gene. The cDNA synthesized in this way was cloned into the prokaryotic vector pET-28a(+), which was then expressed in E. coli cells. The correctness of this approach was confirmed by sequencing resultant cDNA sequencing, which showed full (100%) identity with the nucleotide sequence available in the GenBank database (accession code: GenBankNM_001014167).

Article Details

How to Cite
Fedchenko, V., Kaloshin, A., & Medvedev, A. (2023). Improvement of the Exon Method for Rapid Synthesis of cDNA of the Rat Renalase Gene. Biomedical Chemistry: Research and Methods, 6(3), e00201. https://doi.org/10.18097/BMCRM00201
Issue
Vol. 6 No. 3 (2023)
Section
EXPERIMENTAL RESEARCH

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