Expression vector construction method for Arabidopsis thaliana FIPV promoter fusion GUS gene

• Main Authors: SONG WENLU, DU XINXIN, CHANG YANHONG, DU QIULI, WANG CHAO, YIN CHUNGUANG, XUE LIPING
• Format: Patent
• Language: Chinese; English
• Published: 03.05.2019
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention provides an expression vector construction method for an Arabidopsis thaliana FIPV promoter fusion GUS gene. The arabidopis thaliana at Col-01 week of growing is used as a material to extract the genome DNA (gDNA) by a CTAB method, and the specific primers (F and R) are designed to amplify a FIPV gene promoter, the extracted gDNA is used as a template, the F and R are designed as thespecific primers, subjected to a PCR reaction and then enzyme cutting, and ligated with a Gateway-compatible vector pENTR 3C, the cloning vector pENTR FIPV-Promoter is constructed, and then the pENTR FIPV-Promoter is subjected to a LR reaction with the vector pMDC162 containing the GUS reporter gene, and the expression vector pMDC FIPV-Promoter-GUS of FIPV-Promoter fusion GUS is obtained. 本发明是一种拟南芥FIPV启动子融合GUS基因的表达载体构建方法，以拟南芥Col-01周大的植株为材料，采用CTAB法提取其基因组DNA(gDNA)，设计特异性引物(F和R)扩增FIPV基因启动子，以提取的gDNA为模板，设计的F和R为特异性引物，进行PCR反应再经过酶切，与Gateway兼容的载体pENTR 3C进行连接，构建克隆载体pENTR FIPV-Promoter，然后将pENTR FIPV-Promoter与包含GUS报告基因的载体pMDC162进