METHOD OF PREPARING DNA FRAGMENTS, MONOCLONAL ANTIBODIES 8C1 AND 12G1 INHIBITING ENZYMATIC ACTIVITY OF DNA-POLYMERASE OF THERMUS AQUATICUS AND STRAINS OF HYBRID CULTURED CELLS OF MUS MUSCULUS - PRODUCERS OF MONOCLONAL ANTIBODIES 8C1 AND 12G1

摘要

FIELD: biotechnology, genetic engineering. SUBSTANCE: the parent DNA is incubated with oligonucleotide primer, thermophilic DNA-polymerase and deoxyribonucleoside triphosphates followed by amplification by the repeated temperature cycles. DNA-polymerase is used from Thermus aquaticus cells. Before process onset monoclonal antibodies 8C1 or 12G1 were added to reaction mixture that inhibit enzymatic activity of DNA-polymerase at temperature below 70 C and releasing its after the first temperature cycle termination. The proposed monoclonal antibodies 8C1 were produced by the strain of hybrid cultured cells Mus musculus L. VSSK(P) 643-D of class Ig G2 and has binding constant 1.2 x 10⁹ M. Monoclonal antibodies 12G1 were produced by the strain of hybrid cultured cells Mus musculus L. VSKK(P) 645-D of class Ig G1 and has binding constant 2.7 x 10⁸ M. Strain of hybrid cultured cells Mus musculus L. VSKK(P) 643-D is used as monoclonal antibodies 8C1 producer and the strain of hybrid cultured cells Mus musculus L. VSKK(P) 645-D is used as monoclonal antibodies 12G1 producer also. EFFECT: improved method of preparing. 6 cl, 1 dwg