Multiplex PCR test-system development for the genotyping of transgenic mice with pBC1 MILK expression vector-based transgene

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Abstract


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.


About the authors

V. A. Kalmykov

Institute of Gene Biology of the Russian Academy of Sciences

Email: deikin@igb.ac.ru

Russian Federation, 34/5, Vavilova street, Moscow, 119334

P. A. Kusov

Institute of Gene Biology of the Russian Academy of Sciences; Skolkovo Institute of Science and Technology

Email: deikin@igb.ac.ru

Russian Federation, 34/5, Vavilova street, Moscow, 119334; 3, Nobelya street, Moscow, 143026

A. V. Deykin

Institute of Gene Biology of the Russian Academy of Sciences; Institute of General Pathology and Pathophysiology

Author for correspondence.
Email: deikin@igb.ac.ru

Russian Federation, 34/5, Vavilova street, Moscow, 119334; 8, Baltiyskaya st., Moscow, 125315

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