Construction, Expression, and Purification of ZNF191 (243-368) Zinc Finger Deletion Mutants

Dongxin Zhao ,Zhong-Xian Huang

doi:10.32861/jbr.411.80.82

Abstract

The C-terminal region of the zinc finger protein ZNF191(243-368) contains a putative DNA-binding domain containing four Cys2His2 zinc finger motifs. To understand the properties and functions of the zinc finger motifs, a deletion gene of ZNF191(243-368) was inserted into pGEX-4T-2. The recombinant vector was transformed into Escherichia coli BL21, and a glutathione S-transferase (GST) fusion protein was expressed and purified using glutathione agarose affinity resin. The results show that this expression system can be used to express and purify zinc finger deletion mutants of ZNF191(243-368), providing a basis for further investigation of this protein.

Highlights

Thin films have two important features: (i) thickness of under micron and closer to nanometer; (ii) extremely large surface to thickness of layer
The X-ray diffraction (XRD) and scanning electron microscopy (SEM) and atomic force microscopy (AFM) results confirmed the presence of gold particle in thin films
These films were characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM) and atomic force microscopy (AFM)

Summary

Introduction

Thin films have two important features: (i) thickness of under micron and closer to nanometer; (ii) extremely large surface to thickness of layer. These properties because that ratio surface to bulk is large and are created certain features. Gold in the form of thin films is nowadays used in a vast range of applications such as microelectro mechanical and nanoelectromechanical systems [11, 12], sensors [13], electronic textiles [14], bioengineering [15], generator of nonlinear optical properties [16], or devices for surface-enhanced Raman scattering [17]. Au alloy thin film was created on glass substrate, and various heat treatments was conducted. These films were characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM) and atomic force microscopy (AFM)

Experimental

X-Ray Analysis

SEM Observation

AFM Observation

Conclusions