A modified T-vector for simplified assembly of hairpin RNAi constructs |
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| Authors: | Keming Luo Scott A Harding Chung-Jui Tsai |
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| Institution: | Biotechnology Research Center, School of Forest Resources and Environmental Science, Michigan Technological University, Houghton, MI 49931, USA. |
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| Abstract: | 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. |
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| Keywords: | Cloning vector Dihydroflavonol reductase genes Gene silencing GFP Populus Transgenic plant |
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