A modified T-vector for simplified assembly of hairpin RNAi constructs.
We describe a modified T-vector, pGFPm-T, for direct cloning of RT-PCR products to generate bidirectional restriction fragments for assembly of hairpin-containing RNAi vectors in the popular pFGC and pGSA binary vector backbone. Green fluorescence protein (GFP) is used as a visual reporter for direct selection of recombinants under UV illumination. The simplified cloning process enables a seamless workflow from candidate gene selection and RT-PCR verification to inverted repeat cloning, using a single pair of gene-specific primers.
Luo
Keming
K
Biotechnology Research Center, School of Forest Resources and Environmental Science, Michigan Technological University, Houghton, MI 49931, USA.
Harding
Scott A
SA
Tsai
Chung-Jui
CJ
D000429:Alcohol Oxidoreductases / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics*; D032107:Populus / Q000235:genetics*; D034622:RNA Interference; D018749:RNA, Plant / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*
D010940:Plant Proteins; D018749:RNA, Plant; D049452:Green Fluorescent Proteins; D000429:Alcohol Oxidoreductases; C056570:dihydroflavanol 4-reductase