ABSTRACT
The promoter of mitochondria-localized small heat shock protein gene in Lycopersicon esculentum (Lehsp23.8) is characterized as strongly heat-inducible. In this study, to determine how the expression of Lehsp23.8 is regulated, we conducted five expression vectors carrying the gus gene driven by the 5' deletion products of the Lehsp23.8 promoter. The corresponding transgenic tobacco plants were generated via Agrobacterium tumefaciens-mediated transformation. Transgenic plants were identified by PCR and Southern blotting analysis. GUS activities under heat-shock conditions were characterized in transgenic tobacco plants. After heat shock, obvious GUS staining was detected in the leaves, shoots, roots, flowers and fruits of the transgenic tobacco plants. The result of fluorometric GUS assays in leaves showed that the heat-induced GUS activity of the 565 bp promoter was the strongest, while that of the 255 bp promoter was the lowest. Deletion analysis shows that the smallest promoter fragment (-255 bp to -23 bp) is sufficient for heat induction. It also indicates that the sequences between -255 bp and -565 bp serve as enhancers, while the sequences between -565 bp and -871 bp can repress the heat-induced activity of the Lehsp23.8 promoter.
Subject(s)
Gene Expression Regulation, Plant , Genetics , Genetic Vectors , Heat-Shock Proteins , Genetics , Heat-Shock Proteins, Small , Genetics , Metabolism , Hot Temperature , Solanum lycopersicum , Genetics , Mitochondria , Genetics , Metabolism , Plants, Genetically Modified , Genetics , Promoter Regions, Genetic , Genetics , Nicotiana , GeneticsABSTRACT
AIM: To study the change of blood lipid and difference of expressed gene in chest aorta cells of atherosclerosis model rats with or without the treatment of Taishan Ganoderma Lucidum amylose.METHODS: The concentrations of triglyceride (TG), total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) in serum were measured by automatic serum biochemical assay. Differential-display reverse transcription-PCR (DDRT-PCR) was used to analyze the differential expressed gene in chest aorta cells of atherosclerosis model rat with or without the treatment of Taishan Ganoderma Lucidum amylose. The differential expressed cDNA was confirmed by reverse Northern blotting.RESULTS: Up-regulated and down-regulated cDNA were isolated in chest aorta cells of atherosclerosis model rats with or without Taishan Ganoderma Lucidum amylose treatment. The known up-regulated genes included cytB561, ERP72, nitric oxide synthase 2 and Gpx1. The known down-regulated genes included thymosin β4, FGFRI oncogene, ICAM-1, FK506 binding protein, c-myc and TNF-α. CONCLUSION: Taishan Ganoderma Lucidum amylose decreases the level of blood lipid.