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Cloning and Analysis of SlIAA9 and SlIAA14 Gene Promoters on Tomato

Author: LiXiaoJing
Tutor: ChenYinHuaï¼›ZhangJunHong
School: Hainan University
Course: Crop Genetics and Breeding
Keywords: Tomato SlIAA9 gene SlIAA14 gene Promoter analysis YFP, GUS expression
CLC: S641.2
Type: Master's thesis
Year: 2010
Downloads: 64
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Abstract


Eukaryotic gene expression and regulation of molecular biology research hotspot. Procedures, time and location of gene expression at different levels by different adjustment factors control this complex control mechanism to determine the expression level of the gene and the expression of temporal and spatial order, which is the most important to the regulation on the transcriptional level. In-depth understanding of the structure and regulation of the promoter mode is not only of great theoretical significance, but also has important practical significance to the development of genetic engineering industry. Tomato (Solanum lycopersicum) is a world widely planted vegetables, at the same time, it is also a model plant in plant genetic engineering research, in basic research has been carried out more in-depth genetic transformation system has matured conducive the conduct of this trial. In this study, the use of homologous sequences law and chromosome walking techniques obtained the tomato AUX / IAA family SlIAA9, SlIAA14 gene 5 'end the the upstream PSlIAA9 and PSlIAA14 promoter fragment. Construction of the two fragments and YFP-GUS gene fusion plant the expression vector SlIAA9 :: GUS YFP, YFP as well PSlIAA9 SlIAA14 :: GUS gene driven SlIAA9 overexpression the carrier SlIAA9 :: SlIAA9 and by Agrobacterium tumefaciens in transgenic the method transforming tomato, means the use of transgenic analysis the gene promoter SlIAA9 and SlIAA14 structural features, their expression patterns for further insight into the molecular mechanisms of auxin regulation of tomato growth, revealing the tomato SlIAA9 biology and SlIAA14 gene function and its possible The mechanism of action laid the foundation. The main results obtained in this study are as follows: PCR produces amplification of the gene promoter region of SlIAA9 long of 1891 bp PSlIAA9 sequence, and SlIAA14 gene 2197 bp PSlIAA14 promoter sequence, bioinformatic analysis showed that the obtained sequence fragments ABA, gibberellin, cytokinin hormone action on containing the response and response to abiotic stresses such as drought and damage other acting elements. Building the the promoter fragment PSlIAA9 and PSlIAA14 YFP GUS protein fusion vector SlIAA9 :: GUS YFP, the YFP, PSlIAA9 of SlIAA14 :: GUS driven SlIAA9 gene overexpression carrier SlIAA9 :: 3 SlIAA9. By Agrobacterium mediated transformation of tomato Ailsa Craig the cotyledon get the multiple strains of transgenic plants of three carriers. 4. By stereological fluorescence microscope and GUS staining observed SlIAA9 :: GUS YFP, YFP SliAA14 :: GUS transgenic plants and root YFP and GUS protein expression, found its expression pattern with the apparent specificity, which SlIAA9 :: GUS YFP carrier mainly in the vascular tissue of the leaf base and lateral root primordia parts, while SlIAA14 :: GUS YFP carrier is mainly concentrated in the veins vascular tissue and root tip meristem. 5. Observed SlIAA9 :: SlIAA9 tomato entire mutant phenotype of the transgenic plants, found SlIAA9 gene promoter-driven SlIAA9 gene has not resumed tomato entire mutant single leaf phenotype, its possible causes pending further follow-up studies.

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CLC: > Agricultural Sciences > Gardening > Vegetable gardening > Solanaceous > Tomatoes ( tomatoes )
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