Expression of recombinant major surface protein 5 of Anaplasma marginale (A. marginale) at different temperatures

Authors

  • Amaya Watthanadirek Faculty of Environment and Resource Studies, Mahidol University
  • Thunyaporn Doungdaolek Faculty of Medical Science, Naresuan University
  • Preeyaporn Koedrith Faculty of Environment and Resource Studies, Mahidol University
  • Panat Anuracpreeda Institute of Molecular Biosciences, Mahidol University

Keywords:

Bovine anaplasmosis, Anaplasma marginale, MSP5, recombinant protein, protein expression

Abstract

Background : Bovine anaplasmosis is an important tick-borne disease caused by Anaplasma marginale (A. marginale) and infected in ruminants, mostly in cattle. This disease occurs in tropical and subtropical regions including Thailand and causes a major problem to livestock productions. The major surface protein 5 (MSP5) is one of outer membrane protein of A. marginale which as an immunodominant protein encoded by a single gene and also highly conserved gene.

Objective : The aim of this study was to optimize the conditions for the expression of recombinant major surface protein 5 (rMSP5) of A. marginale.

Methods : The msp5 gene of A. marginale was cloned into the pET100/D-TOPO vector to produce an pET100-msp5-6xHis fusion gene construct. The recombinant proteins were expressed by the plasmids in Escherichia coli host strain BL21 starTM (DE3) at different temperatures (16, 25 and 37 oC) for 6 h. The proteins were analyzed by SDS-PAGE and confirmed the target protein by Western blotting using antisera against His.

Results : After induction with 0.1 mM of Isopropyl gif.latex?\beta-D-1-thiogalactopyranoside (IPTG) at different temperatures for protein expression, the protein was not produced at 6 h for 16 oC. On the other hand, the rMSP5 protein was produced at 25 and 37 oC for 2-6 h but the expressive protein at 25 oC showed lower yield than that at 37 oC.

Conclusion : In this study, the best condition for rMSP5 protein expression was cultured at 37 oC for 4 h. The protein was identified as the rMSP5 at the molecular weight of 26 kDa.

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References

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Published

2023-08-16

How to Cite

1.
Watthanadirek A, Doungdaolek T, Koedrith P, Anuracpreeda P. Expression of recombinant major surface protein 5 of Anaplasma marginale (A. marginale) at different temperatures. Chula Med J [Internet]. 2023 Aug. 16 [cited 2024 Nov. 6];62(3). Available from: https://he05.tci-thaijo.org/index.php/CMJ/article/view/349