tailieunhanh - báo cáo khoa học: " Cloning of transgenic tobacco BY-2 cells; an efficient method to analyse and reduce high natural heterogeneity of transgene expression"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Cloning of transgenic tobacco BY-2 cells; an efficient method to analyse and reduce high natural heterogeneity of transgene expression | BMC Plant Biology BioMed Central Research article Cloning of transgenic tobacco BY-2 cells an efficient method to analyse and reduce high natural heterogeneity of transgene expression Eva Nocarova and Lukas Fischer Open Access Address Charles University in Prague Faculty of Science Department of Plant Physiology Vinicna 5 CZ 128 44 Prague 2 Czech Republic Email Eva Nocarova - evanocarova@ Lukas Fischer - lukasf@ Corresponding author Published 22 April 2009 Received 30 September 2008 BMC Plant Biology 2009 9 44 doi 1471-2229-9-44 Accepted 22 April 2009 This article is available from http 1471-2229 9 44 2009 Nocarova and Fischer licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License http licenses by which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. Abstract Background Phenotypic characterization of transgenic cell lines frequently used in plant biology studies is complicated because transgene expression in individual cells is often heterogeneous and unstable. To identify the sources and to reduce this heterogeneity we transformed tobacco Nicotiana tabacum L. BY-2 cells with a gene encoding green fluorescent protein GFP using Agrobacterium tumefaciens and then introduced a simple cloning procedure to generate cell lines derived from the individual transformed cells. Expression of the transgene was monitored by analysing GFP fluorescence in the cloned lines and also in lines obtained directly after transformation. Results The majority 90 of suspension culture lines derived from calli that were obtained directly from transformation consisted of cells with various levels of GFP fluorescence. In contrast nearly 50 of lines generated by cloning cells from the primary heterogeneous suspensions consisted of cells with homogenous GFP fluorescence. The rest