METHOD FOR DETECTION OF RECOMBINANT GENE OF ACYL-LIPIDIC DESATURASE, JOINED WITH THERMO RESISTANT LICHENASE, IN GENETICALLY MODIFIED PLANT BY MEANS OF MULTIPLEX POLYMERASE CHAIN REACTION

摘要

A method for detection of recombinant gene of acyl-lipidic desaturase, fused with thermoresistant lichenase, in genetically modified plant by means of multiplex polymerase chain reaction, for realization of which the denaturation of plant DNA is conducted; the cycles, each of which comprises DNA denaturation, plant DNA renaturation with oligonucleotide primers, synthesis of fragments of target genes. The pairs of oligonucleotide primers are used for conducting of multiplex polymerase chain reaction, complementary to the recombinant gene of acyl-lipidic desaturase, joined with thermo resistant lichenase, to the genes of actin andsubunit of tobacco GTP-binding protein, to the genes of virDl and virE Agrobacterium tumefaciens, gene nptII and terminator of gene of nopaline synthase, and amplification is realized under following conditions: denaturation of plant DNA during 5 minutes at 94 °C; 30 cycles, each of which comprises DNA denaturation during 30 sec. at 94 °C, renaturation of plant DNA with oligonucleotide primers during 45 sec. at 61 °C , synthesis of fragments of target genes during 45 sec. at 72 °C; synthesis of fragments of target genes during 5 minutes at 72 °C.