Biotechnology Letters

, Volume 30, Issue 7, pp 1271–1274 | Cite as

A modified T-vector for simplified assembly of hairpin RNAi constructs

Original Research Paper


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.


Cloning vector Dihydroflavonol reductase genes Gene silencing GFP Populus Transgenic plant 



We thank Dr. Soon-Yong Choi (HanNam University, South Korea) for providing the pHNT vector. This work was supported by the US Department of Energy, Office of Science, Biological and Environmental Research Program (DE-FG02-05ER64112), and the US National Science Foundation Plant Genome Program (DBI-0421756).


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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  1. 1.Biotechnology Research Center, School of Forest Resources and Environmental ScienceMichigan Technological UniversityHoughtonUSA

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