A novel sequence that functions as a promoter element for moderate constitutive expression of transgenes, designated as the PtMCP promoter, was isolated from the woody perennial Populus *** PtMCP promoter was fused to...
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A novel sequence that functions as a promoter element for moderate constitutive expression of transgenes, designated as the PtMCP promoter, was isolated from the woody perennial Populus *** PtMCP promoter was fused to the GUS reporter gene to characterize its expression pattern in different *** stable Arabidopsis transformants, transcripts of the GUS reporter gene could be detected by RT-PCR in the root, stem, leaf, flower and *** histochemical and fluorometric GUS activity assays demonstrated that the promoter could direct transgene expression in all tissues and organs, including roots, stems, rosette leaves, cauline leaves and flowers of seedlings and maturing *** constitutive expression pattern was similar to that of the CaMV35S promoter,but the level of GUS activity was significantly lower than in CaMV35S promoter∷GUS *** also characterized the promoter through transient expression in transgenic tobacco and observed similar expression *** GUS staining and quantitative analysis detected GUS activity in all tissues and organs of tobacco, including roots, stems, leaves, flower buds and flowers, but GUS activity in PtMCP promoter∷ GUS plants was significantly lower than in CaMV35 S promoter∷ GUS *** results suggested that the PtMCP promoter from poplar is a constitutive promoter with moderate activity and that its function is presumably conserved in different ***, the PtMCP promoter may provide a practical choice to direct moderate level constitutive expression of transgenes and could be a valuable new tool in plant genetic engineering.
TRAP法是一种常用的测定端粒酶活性的方法,但并不太适合测定木本植物的端粒酶活性.因此以油松针叶为实验材料,对TRAP法进行改进.参照前人的研究通过对比实验证明,在端粒酶提取过程中将PEG8000加入上清液,可获得高效的端粒酶.在此基础上,对PCR先导引选择TS、模板浓度选择100 ng,最终获得油松针叶高质量的端粒酶PCR产物.应用SYBR Green I替代银染液对电泳凝胶进行染色,获得条带清晰、重复性好的电泳结果.通过对油松针叶、银杏叶片、胡杨愈伤组织、沙冬青悬浮细胞等4种木本植物实验材料进行测定,证明该改进的技术体系可能是适合于木本植物端粒酶活性测定的稳定的、灵敏度高的可行方法.
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