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Analysis of inheritability and expression profile of single and multi-copy transgene(s) in rice over generations

机译:单代和多代水稻转基因世代的遗传性和表达谱分析

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Transformation of Oryza sativa subsp indica variety Pusa Basmati 1 with Agrobacterium tumefaciens strain LBA4404(pTOK233) carrying genes coding for neomycin phosphotransferase (nptll), beta-glucuronidase (gus) and hygromycin phosphotransferase (hph)under the control of plant-specific promoters (pnos and pCaMV35S) within its T-DNA region produced transgenics with single and multiple copies of T-DNA. Simple Mendelian as well as complex patterns of the inheritance for hygromycin resistance trait wereobserved in R1 and R2 generations. Non-segregating lines selected in R2 generation did not show further segregation of the resistance trait in R3 and R4 generations accompanied by stabilization of integrated transgenes. One of these lines showed the presence of truncated T-DNA in R1 generation. The single copy transgenics showed high stability of expression of gus gene, whereas multi-copy transgenics were prone to silencing up to R3 generation after which no further reduction in gene expression was observed.
机译:根癌农杆菌菌株LBA4404(pTOK233)在植物特异性启动子(pnos)的控制下转化根癌农杆菌菌株LBA4404(pTOK233)来转化稻稻亚种Pusa Basmati 1 (pCaMV35S)和pCaMV35S)在其T-DNA区域内产生具有单拷贝和多拷贝T-DNA的转基因。在R1和R2世代中观察到了简单的孟德尔性状和潮霉素抗性性状遗传的复杂模式。在R2世代中选择的非分离系没有显示出R3和R4世代中抗性性状的进一步分离,并伴随着整合的转基因的稳定。这些品系之一显示R1世代中存在截短的T-DNA。单拷贝转基因显示了gus基因表达的高度稳定性,而多拷贝转基因则倾向于沉默直至R3世代,此后未观察到基因表达的进一步降低。

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