Cloning, Expression, Purification and Biological Activities of Recombinant Mouse Interleukin-2 in E. coli M15

Authors

  • Sanchai Chantajorn Centre for Agricultural Biotechnology, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand.
  • Ratchanee Hongprayoon Department of Plant Pathology, Faculty of Agriculture at Kamphaeng Saen, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom, 73140, Thailand.
  • Thaweesak Songserm Department of Veterinary Pathology, Faculty of Veterinary, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand.

Keywords:

mouse interleukin-2, cloning, protein expression, protein purification, immunofluorescence

Abstract

Molecular cloning, sequencing and expression of recombinant mouse interleukin 2 (rmIL-2) were described. The interleukin-2 (IL-2) cDNA, 450 base pairs in length with repeating CAG, was amplified using specific primers. The IL-2 cDNA showed high homology at 100%, 100%, 91%, 96% and 94% with five strains of mice previously reported (GeneBank accession number AY147902.1, MMU41494, MMU41504, MMU41505 and MMU41506). The IL-2 gene was cloned into the pDrive cloning vector and consequently expressed using pQE30 expression vector which provided high expression level of the recombinant protein. The predicted rmIL-2 sequence is 161 amino acids with a molecular weight of 19 kDa. The expressed protein was then purified by Ni-NTA column under denaturing condition. Analysis of the rmIL-2 by SDS-PAGE demonstrated two bands of 19 and 38 kDa representing monomeric and dimeric forms of this protein. The biological activity in stimulating T-cell proliferation was also described and the binding signal to the receptor was easily observed by immunofluorescence.

Published

2007-06-30

How to Cite

Sanchai Chantajorn, Ratchanee Hongprayoon, and Thaweesak Songserm. 2007. “Cloning, Expression, Purification and Biological Activities of Recombinant Mouse Interleukin-2 in E. Coli M15”. Agriculture and Natural Resources 41 (2). Bangkok, Thailand:335-45. https://li01.tci-thaijo.org/index.php/anres/article/view/244251.

Issue

Vol. 41 No. 2 (2007): April-June

Section

Research Article

License

online 2452-316X print 2468-1458/Copyright © 2022. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/),
production and hosting by Kasetsart University of Research and Development Institute on behalf of Kasetsart University.