Functionally active rat S100A4 from a polymerase chain reaction‑synthesized gene expressed in soluble form in Escherichia coli

Liu,Ziquan; Xu,Chuanxiang; Zhang,Jianwei; Chen,Yunyun; Liu,Xiaohua; Wu,Lei; Zhang,Zhiqing; Meng,Xiangyan; Liu,Hongtao; Jiang,Zifeng; Wang,Tianhui

doi:10.3892/ol.2014.1870

2014-April Volume 7 Issue 4

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2014-April Volume 7 Issue 4

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Article

Functionally active rat S100A4 from a polymerase chain reaction‑synthesized gene expressed in soluble form in Escherichia coli

Authors:
- Ziquan Liu
- Chuanxiang Xu
- Jianwei Zhang
- Yunyun Chen
- Xiaohua Liu
- Lei Wu
- Zhiqing Zhang
- Xiangyan Meng
- Hongtao Liu
- Zifeng Jiang
- Tianhui Wang
View Affiliations / Copyright

Affiliations: Institute of Health and Environmental Medicine, Heping, Tianjin 300050, P.R. China, Institute of Zoology, Chinese Academy of Sciences, Chaoyang, Beijing 100101, P.R. China
Pages: 1179-1184
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Published online on: February 11, 2014

https://doi.org/10.3892/ol.2014.1870
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Abstract

S100A4 protein is associated with Ca2+‑dependent regulation of intracellular activities and is significant in the invasion, growth and metastasis of cancer. In order to express rat S100A4 functionally and identify its biological activity following purification, an S100A4 gene fragment was optimized and fully synthesized via overlapping polymerase chain reaction. The gene was inserted into the prokaryotic expression vector, pBV220, with phage λ PRPL promoters following confirmation by DNA sequencing. The pBV220‑S100A4 plasmid was constructed and transformed into Escherichia coli DH5α. Following temperature induction, rat S100A4 was overexpressed and the protein was observed to be located in the supernatant of the lysates, which was ~30‑40% of the total protein within the host. The protein was isolated and purified by metal‑chelate affinity chromatography. High purity protein (>98% purity) was obtained and in vitro western blot analysis identified that the recombinant S100A4 was able to bind to the antibody against wild‑type S100A4. The bioactivity of the recombinant protein was detected via Transwell migration and invasion assays. The polyclonal antibody of rat S100A4 protein was prepared for rabbit immunization and exhibited similar efficacies when compared with commercial S100A4. Therefore, rat S100A4 was functionally expressed in E. coli; thus, the production of active recombinant S100A4 protein in E. coli may further aid with the investigation and application of S100A4.

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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Functionally active rat S100A4 from a polymerase chain reaction‑synthesized gene expressed in soluble form in Escherichia coli

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Abstract

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