Expression and purification of recombinant proteins based on human prostate stem cell antigen and heat shock protein-70

  • Authors:
    • Lei Dong
    • Xiaopeng Zhang
    • Changming Yu
    • Jun Ren
    • Lihua Hou
    • Ling Fu
    • Shaoqiong Yi
    • Wei Chen
  • View Affiliations

  • Published online on: February 20, 2013     https://doi.org/10.3892/etm.2013.967
  • Pages: 1161-1164
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Abstract

The aim of this study was to express and purify recombinant proteins based on human prostate stem cell antigen (PSCA) and heat shock protein-70 (HSP70). The PSCA gene and various structural domains of HSP70 were amplified by polymerase chain reaction (PCR) with the respective primers. Then, the PSCA was cloned into the prokaryotic expression vector pET21a(+) with the amino-terminus, carboxyl-terminus and overall length of HSP70, by enzyme digestion to construct the recombinant plasmids pET21-PSCA-HSPN, pET21-PSCA-HSPC and pET21-PSCA-HSP, respectively. After being expressed in Escherichia coli (E. coli) by isopropyl β-D-1-thiogalactopyranoside (IPTG) induction, recombinant fusion proteins were purified. Western blotting was performed to confirm the expression of the recombinant proteins. The results revealed that recombinant plasmids were successfully constructed. The PSCA-HSPC and PSCA-HSP expressed in E. coli existed in soluble form, as confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The purity of the recombinant proteins PSCA‑HSPC and PSCA-HSP reached >95% following purification with the nickel-nitrilotriacetic acid (Ni-NTA) resin, Phenyl-Sepharose Fast Flow and Superdex 75, which lays a foundation for the development of vaccines for prostate cancer.
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April 2013
Volume 5 Issue 4

Print ISSN: 1792-0981
Online ISSN:1792-1015

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Spandidos Publications style
Dong L, Zhang X, Yu C, Ren J, Hou L, Fu L, Yi S and Chen W: Expression and purification of recombinant proteins based on human prostate stem cell antigen and heat shock protein-70. Exp Ther Med 5: 1161-1164, 2013
APA
Dong, L., Zhang, X., Yu, C., Ren, J., Hou, L., Fu, L. ... Chen, W. (2013). Expression and purification of recombinant proteins based on human prostate stem cell antigen and heat shock protein-70. Experimental and Therapeutic Medicine, 5, 1161-1164. https://doi.org/10.3892/etm.2013.967
MLA
Dong, L., Zhang, X., Yu, C., Ren, J., Hou, L., Fu, L., Yi, S., Chen, W."Expression and purification of recombinant proteins based on human prostate stem cell antigen and heat shock protein-70". Experimental and Therapeutic Medicine 5.4 (2013): 1161-1164.
Chicago
Dong, L., Zhang, X., Yu, C., Ren, J., Hou, L., Fu, L., Yi, S., Chen, W."Expression and purification of recombinant proteins based on human prostate stem cell antigen and heat shock protein-70". Experimental and Therapeutic Medicine 5, no. 4 (2013): 1161-1164. https://doi.org/10.3892/etm.2013.967