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Arabidopsis function of unknown genes AtBN3

Author: LiWenJuan
Tutor: ZhaoZhiGuang
School: Lanzhou University
Course: Botany
Keywords: Semi- quantitative RT-PCR Resistance Brassinolide Nitric oxide
CLC: Q943.2
Type: Master's thesis
Year: 2009
Downloads: 22
Quote: 0
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Abstract


Arabidopsis thaliana is the first complete genome sequencing of the higher plant, but so far, there are still a large number of gene function is to be us to explore. With the development of reverse genetics and modern molecular biology technology, these features unknown gene mapping, gene function analysis, gene expression and regulation of research has increasingly gained attention. In this study, genetic engineering, molecular biology and other means to Arabidopsis genome genes of unknown function AtBN3 (by BR-NO regulate gene). AtBN3 located on chromosome IV of Arabidopsis, full-length 2071bp, gene inside without introns. Protein encoded by the gene sequences of the 652 amino acids. Bioinformatics analysis revealed that the genes may respond to various Stress response, but also for a variety of plant hormones BR signal is sensitive. Meanwhile, the promoter cis-regulatory element contains multiple stress responses related components. This thesis studied through experimental methods of molecular biology of the gene in the expression of various stress conditions, BR and NO also studied the expression of this gene. First by semi-quantitative RT-PCR method to detect AtBN3 induced expression in a variety of changes in the results. The results showed that the gene in the hot, cold, salt and osmotic stress, the gene expression level compared with the control has a significant change, and in the synthesis of brassinosteroid mutants defective det2, this change becomes significant difference small. Therefore, we hypothesized that AtBN3 Arabidopsis gene is a gene associated with resilience, and the expression of it in the face of adversity BR entirely by endogenous levels of regulation. Do we have the wild-type Arabidopsis Col-0, brassinosteroid synthesis mutants defective det2 and brassinosteroid signaling mutants bril-5 three kinds of plants as materials, BR, NO donor GSNO and NO scavenger cPTIO different combinations of these three reagents handles these three materials. The results showed that, AtBN3 NO and BR is a common regulation of the gene. BR and NO on AtBN3 have a positive regulatory role, but the processing time associated with. BR endogenous deletion or NO, the gene expression will have an impact. BR induced by this gene levels of endogenous NO; NO induction of this gene is also affected by endogenous BR levels. We also speculate AtBN3 may be located in BR pipeline downstream of a regulatory factor receptor. We semi-quantitative RT-PCR method to detect AtBN3 tissue-specific expression. The results show that its highest expression in the stems, roots, followed by leaves and flowers in the least. Gateway technology constructed with three different vectors: for the study of tissue specificity AtBN3 pBN3 :: GUS promoter fusion expression vector, AtBN3-GFP expression vectors subcellular localization, and ox-AtBN3 overexpression vector. This experiment further study AtBN3 the biological function of the foundation.

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CLC: > Biological Sciences > Botany > Plant Cell Genetics > Plant Genetic Engineering
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