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

Main Article Content

V.I. Fedchenko
A.A. Kaloshin
A.E. Medvedev


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.


  1. An, X., Lu, J., Huang, J.D., Zhang, X., Chen, J., Zhou, Y., Tong, Y. (2007) Rapid assembly of multiple-exon cDNA directly from genomic DNA. PLoSOne, 2, e1179. DOI
  2. Davies W.L., Carvalho L.S., Hunt D.M. (2007) SPLICE: A technique for generation in vitro spliced coding sequeces from genomic DNA. BioTechniques, 43, 785–789. DOI
  3. Jayakumar, A., Huang, W.-Y., Raetz, B., Chirala, S.S., Wakil, S.J. (1996) Cloning and expression of the multifunctional human fatty acid synthase and its subdomains in Escherichia coli. Proc. Natl. Acad. Sci. USA, 93, 14509–14514. DOI
  4. Booth, P.M., Buchman, G.W., Rashtchian, A. (1994) Assembly and cloning of coding sequences for neurotrophic factors directly from genomic DNA using polymerase chain reaction and uracil DNA glycosylase. Gene, 146, 303–308. DOI
  5. Fedchenko, V.I., Kaloshin, A.A., Mezhevikina, L.M., Buneeva, O.A., Medvedev, A.E. (2013) Construction of the Coding Sequence of the Transcription Variant 2 of the Human Renalase Gene and Its Expression in the Prokaryotic System.Int. J. Mol. Sci. v.14, 12764-12779; DOI
  6. Fedchenko, V.I., Kaloshin, A.A. (2019) A simplified method for obtaining cDNA of low-copy and silent eukaryotic genes using human renalase as an the example. Biomedical Chemistry: Research and Methods, 2(2), 1-7. DOI
  7. Fedchenko, V.I., Kaloshin, A.A., Kaloshina, S.A., Medvedev, A.E. (2021). Expression and Isolation of N-Terminal Truncated Human Recombinant Renalase in Prokaryotic Cells. Biomedical Chemistry: Research and Methods, 4(3), e00158. DOI
  8. Tersus Plus PCR kit. Retrived June 01, 2023 from