Expression of recombinant human granulocyte-colony stimulating factor in Escherichia coli using various induction methods

Author:

Dewi K S,Permadi D G,Aminah ,Fuad A M

Abstract

Abstract Granulocyte-colony stimulating factor (G-CSF) is a glycoprotein that has several therapeutic applications. It consists of 174 amino acids and manufactured by recombinant DNA technology. Until now, the Escherichia coli expression system is still become the first choice for producing recombinant proteins. It is because of this organism is simple to culture in low-cost medium and easy to scale up. In the course to find the most efficient way to produce a high yield of recombinant human G-CSF, we compared several types of medium with different induction methods. In this experiment, recombinant E. coli NiCo21(DE3) harbouring gene encoding rh-GCSF proteins were cultured in various media including auto-induction, non-induction, and IPTG-induction. To determine the protein expression profile, culture sampling was done every 12 h (up to 60 h). Then, the optical density at ʎ 600 nm was measured using UV spectrophotometer and rh-GCSF protein expression were characterized using SDS-PAGE and western blot analyses. ImageJ software was used to calculate the amount of rh-GCSF protein yield using Bovine Serum Albumin (BSA) with known concentration as a standard. Result of this experiment concluded that simple auto-induction medium from Imperial College could produce good amount of rh-GCSF proteins (117 µg/mL) with relatively low production cost and short incubation time.

Publisher

IOP Publishing

Subject

General Engineering

Cited by 1 articles. 订阅此论文施引文献 订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3