Open AccessMultiplex PCR test-system development for the genotyping of transgenic mice with pBC1 MILK expression vector-based transgene
Author(s) -
Vladislav Kalmykov,
Vladislav Kalmykov,
Pavel A. Kusov,
Pavel A. Kusov,
Alexey V. Deykin,
А. В. Дейкин
Publication year - 2019
Publication title -
doklady akademii nauk. rossijskaâ akademiâ nauk
Language(s) - English
Resource type - Journals
ISSN - 0869-5652
DOI - 10.31857/s0869-56524856750-753
Subject(s) - transgene , genotyping , biology , microinjection , microbiology and biotechnology , transgenesis , expression vector , heat shock protein , genetics , gene , recombinant dna , genotype , reproductive biology , embryogenesis
Developed multiplex PCR test-system for identification of transgenic cis-regulatory elements (promoter, insulator) from genetic constructions based on pBC1 Milk expression vector incorporated into the Mus musculus genome. Test-system was validated by genotyping producent strains generated by linearized vector DNA microinjection into fertilized zygote with occasional transgene genome integration, expressing the following transgenes in mammary gland, in milk: Human lactoferrin protein in different length, heat-shock protein HSP-70, Firefly luciferase protein, lysozyme protein. Reaction conditions selection described in details for better reproducibility.