ectopic expression of GA 2-oxidase 6 from Brassica napus L. causes dwarfism, late flowering and enhanced chlorophyll accumulation in Arabidopsis thaliana
full-length coding sequence of GA2ox6 amplified by PCR, cloned into pET32a to be expressed as fusion with thioredoxin in Escherichia coli BL21(DE3) for in vitro enzyme activity measurements
functional overexpression in Brassica napus cv. N529 using the Agrobacterium tumefaciens strain GV3101 for transfection, the recombinant expression leads to decreased biomass accumulation and lignification, quantitative real-time PCR expression analysis. The transgenic plants show growth retardation, flowering delay, and dwarf stature, phenotype, overview
PCR-amplifiction of cDNA, inserted via pGEM-T Easy cloning vector and pAHC18 into pCAMBIA1301 and transferred into Agrobacterium tumefaciens (strain EHA105) and used to transform rice and tobacco (Nicotiana tabacum), or for activity assays subcloned into pGEX-5X expression vector to transform Escherichia coli BL21(DE3)
rapid amplification of cDNA ends, expression vector produced by PCR, inserted into PMD 18 T-Simple Vector, inserted into pMAL-c2x vector, transformed and expressed in Escherichia coli Tb1
reporter plasmid: promoter region of GA2ox7 amplified from genomic DNA by PCR, fused to luciferase gene in pUC19, effector plasmid: replacing GUS gene of pBI221 with DDF1 coding region, reference plasmid with Renilla luciferase gene, introduction of the plasmids to leaves via particle bombardment, leaves incubated on 0.8% agar plates at 22°C for 24 h. transgenic plants: PCR-amplificatoin of DDF1 coding region, cloned into pENTER-TOPO, inserted into vector pGWB8, Agrobacterium (strain EHA105)-mediated transformation of wild-type and GA2ox7-2 plants
SLN gene, 2 cDNAs encoding gibberellin 2-oxidases, PsGA2ox1 isolated by screening of a Lambda-ZAP cDNA library, excised into phagemid form and expressed in Escherichia coli, PsGA2ox2 obtained as PCR product and expressed in Escherichia coli
1.14.11.13 gibberellin 2beta-dioxygenase
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