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Extraction and Purification of Chromatin Assembly Associated Proteins and Heat Shock Induced in Vitro Transcription of hsp90α Gene on Chromatin Template

Author: DaiHui
Tutor: JinYuHuai；ZhangYe
School: Hebei Medical University
Course: Pathogen Biology
Keywords: Histone NAP-1 Whole cell extract In vitro transcription RT-PCR Chromatin assembly Histone methylation
CLC: R346
Type: Master's thesis
Year: 2005
Downloads: 60
Quote: 0
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Abstract


Objective: genomic DNA of eukaryotic cells contained in the chromatin of DNA around the histone octamer form nucleosome is the basic unit of chromatin . The formation of chromatin in addition to the packaging DNA , but also by the adjustment of its structure , i.e. groups of protein covalent modification and the ATP-dependent chromatin remodeling complex , effectively \DNA binding , in order to promote or inhibit the expression of genes . Core histone tails occurred many covalent modifications include acetylation , phosphorylation , methylation , ubiquitination , SUMO , etc. , involved in the activation and inhibition of gene regulation process , the formation of the histone code has become an important table The concept of genetic markers . Through the establishment of a chromatin assembled in vitro system , for better study histone modifications and various modifications interaction between the role of the regulation of gene transcription and laid a foundation. In the the Chromatin assembly process ( H3/H4 ) 2 tetramer first DNA binding , followed H2A/H2B two heterodimer binding to (H3/H4) 2 binding at the 5 'and 3'DNA on form nucleosomes body . The study shows that many molecular chaperone involved in nuclear body formation , most of the binding proteins of the core histones , nucleosome assembly protein -1 (nucleosome assembly protein-1, NAP-1) may be first combined with the histone H2A and H2B assist histone and DNA binding . Accordingly, we extracted from HeLa cells purified core histone proteins , and application to the expression of insect expression system , extract chromatin assembly the related factors NAP - 1 , and thus provide effective methods for further studies of gene expression regulation mechanisms in the chromatin templates .

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