Abstract
The routine generation of transgenic plants involves analysis of transgene integration into the host genome by means of Southern blotting. However, this technique cannot distinguish between uniformly transformed tissues and the presence of a mixture of transgenic and non-transgenic cells in the same tissue. On the other hand, the use of reporter genes often fails to accurately detect chimerical tissues because their expression can be affected by several factors, including gene silencing and plant development. So, new approaches based on the quantification of the amount of the transgene are needed urgently. We show here that chimeras are a very frequent phenomenon observed after regenerating transgenic plants. Spatial and temporal analyses of transformed tobacco and apricot plants with a quantitative, real-time PCR amplification of the neomycin phosphotransferase (nptII) transgene as well as of an internal control (beta-actin), used to normalise the amount of target DNA at each reaction, allowed detection of chimeras at unexpected rates. The amount of the nptII transgene differed greatly along with the sub-cultivation period of these plants and was dependent on the localisation of the analysed leaves; being higher in roots and b...Continue Reading
References
Jan 1, 1990·Molecular & General Genetics : MGG·G VancanneytM Rocha-Sosa
Aug 1, 1989·Trends in Genetics : TIG·S Poethig
Feb 1, 1993·Plant Molecular Biology·T Schmülling, J Schell
Oct 1, 1996·Genome Research·C A HeidP M Williams
Jun 13, 2001·Analytical Biochemistry·N Billinton, A W Knight
Jul 24, 2001·BioTechniques·D J InghamG Hansen
Feb 16, 2002·Methods : a Companion to Methods in Enzymology·K J Livak, T D Schmittgen
Oct 26, 2002·BMC Biotechnology·Giovanna MasonGian Paolo Accotto
May 4, 2005·Transgenic Research·César Petri, Lorenzo Burgos
May 17, 2005·Experimental and Molecular Pathology·Shu-Ching Shih, Lois E H Smith
Dec 14, 2006·Plant Cell Reports·Elena Rakosy-TicanMichael R Davey
Dec 15, 2006·Biotechnology Journal·Behrooz DarbaniWilliam N Camargo
Jul 12, 2007·Plant Cell Reports·Xue-Yun ZhuLi-Ping Chen
Dec 6, 2007·BMC Bioinformatics·Joshua S YuanC Neal Stewart
Dec 15, 2007·Analytical Biochemistry·Cheng Xin YiYan Hong
May 2, 2008·Plant Cell Reports·César PetriLorenzo Burgos
Feb 6, 2009·Biotechnology Progress·Ahmad A OmarJude W Grosser
Jan 1, 1995·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·J CooleyP Christou
Jul 1, 2001·Plant Cell Reports·M Schmidt, W Parrott
Citations
Jul 21, 2010·Plant Cell Reports·I M G Padilla, L Burgos
Jun 21, 2011·Yi chuan = Hereditas·Yan WangLi-Ping Chen
Nov 15, 2011·Biotechnology Advances·Biao WangQiaochun Wang
Dec 26, 2012·Gene·Shweta Mehrotra, Vinod Goyal
Jul 12, 2014·Transgenic Research·Julia RubioHumberto Prieto
Aug 7, 2016·Frontiers in Plant Science·Nathaniel M ButlerDavid S Douches
Aug 29, 2016·Transgenic Research·Karin FisterBorut Bohanec
Aug 30, 2017·Plant Cell Reports·Yue Sun, Priya Aiyar Joyce
Apr 2, 2020·PloS One·Mihály KondrákZsófia Bánfalvi
Apr 14, 2018·Transgenic Research·Cesar PetriChris Dardick
May 14, 2020·BMC Biotechnology·Zsófia BánfalviMihály Kondrák
Mar 12, 2020·Physiology and Molecular Biology of Plants : an International Journal of Functional Plant Biology·Amanpreet KaurAnil Kumar
Dec 13, 2019·Royal Society Open Science·Shengxing LiHanyao Zhang