Method for identifying copy number of T-DAN tandem repeat sequences in transgenic plant through real-time fluorescence quantification PCR method

The invention discloses a method for identifying the copy number of T-DAN tandem repeat sequences in a transgenic plant through a real-time fluorescence quantification PCR method. The method comprises the following steps: connecting an HMGA gene to a plant expression vector Pski015 containing a scre...

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Bibliographische Detailangaben
Hauptverfasser:	LIU JINGJING, WEI SHU, XI YUZHEN, SONG DAPENG
Format:	Patent
Sprache:	eng
Schlagworte:	AGRICULTURE ANIMAL HUSBANDRY BEER BIOCHEMISTRY CHEMISTRY COMPOSITIONS OR TEST PAPERS THEREFOR COMPOSITIONS THEREOF CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES CULTURE MEDIA ENZYMOLOGY FISHING FORESTRY HUMAN NECESSITIES HUNTING MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING NEW PLANTS OR PROCESSES FOR OBTAINING THEM PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES PROCESSES OF PREPARING SUCH COMPOSITIONS PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS TRAPPING VINEGAR WINE
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Beschreibung
Zusammenfassung:	The invention discloses a method for identifying the copy number of T-DAN tandem repeat sequences in a transgenic plant through a real-time fluorescence quantification PCR method. The method comprises the following steps: connecting an HMGA gene to a plant expression vector Pski015 containing a screening gene BAR through a Southern Buddhism mediated plant transformation method; then transfecting wild Col-4 arabidopsis; connecting the vector with two reference plasmids; extracting RNA of glyphosate-screened transgenic arabidopsis and carrying out inverse transcription to obtain cDNA as a detection template; and detecting the copy number of a mutant plant in a way of real-time quantification PCR respectively by virtue of quantitative primers of a glyphosate-resistance gene and the HMGA gene. The method disclosed by the invention has the beneficial effects that the influence of a T-DAN tandem structure on identification of the copy number of a transgenic plant can be effectively shielded; and compared with Southern blot, quantification PCR is more convenient and more efficient.