Development of a Multiplex PCR Test System for the Determination of a Transgene Based on the pBC1 Plasmid and Its Derivatives for the Expression of Recombinant Proteins in Mus musculus Milk.

Abstract

A multiplex PCR test system for identification of the regulatory sequences of genetic constructs for transformation (promotor, insulator, and terminator) in the Mus musculus genome and for transgenic animal selection by genotyping with horizontal agarose gel electrophoresis detection was developed. The proposed system was validated by genotyping mouse strains producing human lactoferrin, heat shock protein HSP 70, firefly luciferase, and lysozyme, which were obtained by microinjections of linearized DNA into murine zygote pronucleus with random transgene integration into the genome using the pBC1 plasmid for expression of the gene of interest in milk of transformed animals (milk expression vector kit).