Improvement of the Exon Method for Rapid Synthesis of cDNA of the Rat Renalase Gene-Reference-Cited by-同舟云学术

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

Published:2023 Issue:3 Volume:6 Page:e00201
ISSN:2618-7531
Container-title:Biomedical Chemistry: Research and Methods
language:en
Short-container-title:BMCRM

Author:

Fedchenko V.I.¹,Kaloshin A.A.¹,Medvedev A.E.¹

Affiliation:

1. Institute of Biomedical Chemistry, Moscow, 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).

Publisher

Institute of Biochemistry

Subject

General Medicine

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