Dissertation > Excellent graduate degree dissertation topics show

A Preliminary Study on a Putative Plant Transcription Factors Family

Author: WangHongGang
Tutor: ChenLiang
School: Xiamen University
Course: Cell Biology
Keywords: Transcription factor Bioinformatics The prokaryotic expression Subcellular localization
CLC: Q943
Type: Master's thesis
Year: 2008
Downloads: 158
Quote: 1
Read: Download Dissertation

Abstract

Transcription factors (transcription factor, TF) is a class of the same eukaryotic promoter specific DNA sequence binding protein, plays an important role in the regulation of plant growth and development in response to the external environment. Through bioinformatic analysis inferred gene family of transcription factors for the new plant, we carried out a preliminary study, as follows: (1) through bioinformatics databases and related software, we studied transcription factor family the bioinformatics analysis. Found that the multiple sequence alignment and phylogenetic analysis of these members are divided into four subfamilies; EST databases to retrieve information on the expression of these genes found in the family transcription factor family gene members including two conservative district; most of the Arabidopsis gene expression in flowers, seeds, and most of the rice gene expression in flowers, buds and callus; predicted secondary structure, these proteins contain zinc finger structure, low complexity regions, DUF1644 ring finger structure of four types of structural domains, indicating that the function of the family members may have a transcription factor. (2) The Arabidopsis genome DNA the experimental material cloning genes At1g15430, At5g38370, rice genome DNA the cloned gene 0s09g27860 experimental material, 0s01g42700, 0s02g35840 then build its pGEX-4T-2 prokaryotic expression vector, into the expression strain colon Bacillus BL21 (DE3) and Rosetta ^{TM prokaryotic expression of the target protein exists mainly in the form of inclusion bodies. Optimal induction conditions: the induction cell density OD ₆₀₀ of about 0.6 induced matter IPTG concentration of 1.0 mmol / L, the induced temperature 28.5 ° C, induction time of 4 h. GST Resin separation and using a small amount of soluble fusion protein GST-At1g15430. (3) the use of bypass PCR approach the cloned chimeric gene At1g15430-GFP, At5g38370-GFP, 0s01g42700-GFP, 0s09g27860-GFP, and the chimeric gene fragment was ligated into the cloning vector pMD18-T. After sequencing, the use of restriction endonucleases to cut from pMD18-T, expression vector construction pBPF. Through gene gun transformation method, its subcellular localization studies into onion epidermal cells. Onion epidermal cells 0s01g42700-GFP in the nucleus and cytoplasm are green fluorescent by the Bio-RadMRC1024 confocal laser scanning microscope, turn, turn At5g38370-GFP onion epidermal cells with green fluorescence in the nucleus to turn At1g15430-GFP onion epidermal cells with green fluorescence in the cytoplasm turn 0s09g27860-GFP onion epidermal cells was not observed green fluorescence. The results show that of 0s01g42700 a GFP gene expression product parts are transported into the nucleus of all At5g38370-GFP gene expression product transport into the nucleus, At1g15430-GFP gene expression product transport into the nucleus 0s09g27860-GFP genes expressed.}

A Preliminary Study on a Putative Plant Transcription Factors Family

Abstract

Related Dissertations