Expression of a Fusion Protein of Human Proinsulin with Glutathione-S-Transferase in Escherichia coli

Zhi Xin Di; Jian Zhong Ma; Yong Gang Wang

doi:10.4028/www.scientific.net/AMR.998-999.248

Paper Titles

Effects of Echinomycin on PCNA-Dependent Follicular Development in PMSG-Induced Sprague-Dawley Rats
p.228

Effects of Immediate Stimulation on Single Nucleotide and Catecholamine Level in Chinese Tree Shrews
p.233

Effects of Self-Assembling Peptide RADA16-І Hydrogel on Neural Differentiation of Bone Marrow Mesenchymal Stem Cells
p.238

Exercise Induce HO-1 Expression in Skeletal Muscle and Improve Sports Ability of Mice
p.243

Expression of a Fusion Protein of Human Proinsulin with Glutathione-S-Transferase in Escherichia coli
p.248

Foxp3 in Lewis Lung Cancer Cells Enhances the Expression of TGF-β1 and IL-10
p.252

In Situ Imaging of Cancer Cell via Cancer Aptamer and its Fluorescent Clusters
p.256

Inhibition of HIF-1α Signaling in the Ovaries of Sprague-Dawley Rats with Polycystic Ovary Syndrome
p.260

IRE1a Negative Regulates Chondrocyte Differentiation
p.265

HomeAdvanced Materials ResearchAdvanced Materials Research Vols. 998-999Expression of a Fusion Protein of Human Proinsulin...

Expression of a Fusion Protein of Human Proinsulin with Glutathione-S-Transferase in Escherichia coli

Abstract:

A DNA sequence encoding for the human proinsulin was designed according to the codon bias of Escherichia coli and then chemically synthesized. The synthesized DNA fragment was subcloned into pGEX-3X for expression in E. coli BL21 (DE3) and E. coli BL21 Star (DE3), respectively. Conditions for the highest expression of the GST-proinsulin fusion proteins were optimized. These conditions are that cells of E. coli BL21 star (DE3) are incubated in 100mL of the LB medium with 2 mmol/L IPTG and 60μg/mL ampicillin at 26^oCfor 4h. After disrupted E. coli cells with ultrasonication, inclusion bodies were precipitated from cell lysis and washed. Fusion proteins from the inclusion bodies were redissolved in 8mmol/L of urea. After dialysed in purified water, fusion proteins were analysed by SDS-PAGE. The purity of the fusion protein is about 80.5% in total. The fusion protein from SDS-PAGE was further identified by mass/mass spectrum. GST in the dyad protein is confirmed by the 9 matched sequences. However, the left part is proved a polypeptide of which is completely different from the human proinsulin.

You might also be interested in these eBooks

Advances in Applied Sciences, Engineering and Technology II

View Preview

Info:

Periodical:

Advanced Materials Research (Volumes 998-999)

Pages:

248-251

DOI:

https://doi.org/10.4028/www.scientific.net/AMR.998-999.248

Citation:

Cite this paper

Online since:

July 2014

Authors:

Zhi Xin Di, Jian Zhong Ma*, Yong Gang Wang

Keywords:

Fusion Protein, Glutathione-S-Transferase, Human Proinsulin, Mass Spectrum

Export:

RIS, BibTeX

Price:

Permissions CCC:

Request Permissions

Permissions PLS:

Request Permissions

Сopyright:

Citation:

* - Corresponding Author

References

[1] Aldana Trabucchi, Luciano Lucas Guerra, Natalia Inés Faccinetti, Ruben Francisco Iacono, Edgardo Poskus, Silvina Noemí Valdez. Appl Microbiol Biotechnol . 94 (2012), p.1565.

DOI: 10.1007/s00253-011-3721-5

Google Scholar

[2] Youshang Zhang: In Chinese. Food and Drug. 10(2008), p.1.

Google Scholar

[3] Yan Chen, Jinzhao Li, Lingzhao Xia, Ying Zhang, Qiang Li: In Chinese. Chinese Biochemical Journal. 11(1995), p.636.

Google Scholar

[4] Bing Hu, Chao Huang, Wenjing Li, Liuhong Deng, Susheng Xiao, Chunfa Zhang: In Chinese. Biotechnology Bulletin. (2009), p.96.

Google Scholar

[5] Jinqiu Chen, Hongtao Zhang, Meihao Hu, Jianguo Tang. Applied Biochemistry and Biotechnology. 55(1995), p.5.

Google Scholar

[6] Joseph Sambrook, Davide w. Russell, in: Molecular Cloning A laboratory Manual, 3rd, chapter, 15, Kluwer Academic Publishers (2002).

Google Scholar