| 摘要 |
FIELD: medicine, pharmaceutics.SUBSTANCE: invention relates to genetic engineering and may be used in biotechnology to produce proteins of practical interest, which are a product of silent genes or low expression genes. What is presented is a method for producing a recombinant protein of human renalase 2 involving a) preparing a complete cDNA sequence, and b) expressing the prepared sequence in E.coli, wherein the stage (a) is performed by PCR amplification of each exon using specific primer pairs and a genomic DNA as a matrix, followed by twinning requiring no pre-treatment and involving the same method for adjacent exons at first, and then being the product of their amplicon combining. In this case, the primers twice exceeding exons in number are selected so that the forward primer contain a terminal sequence of the preceding exon and an initial sequence of the exon to be copied; the downstream primers contain only a sequence complementary to the end of the amplified exons, while the forward and downstream primers flanking the complete coding sequence contain restriction sites necessary for integration into a plasmid vector.EFFECT: preparing the proteins of practical interest which are the products of silent genes or low expression genes.2 tbl, 7 dwg, 1 ex |
