•  
  •  
 
Chulalongkorn Medical Journal

Abstract

Background : Bovine anaplasmosis is an important tick-borne disease caused byAnaplasma marginale (A. marginale) and infected in ruminants, mostly incattle. This disease occurs in tropical and subtropical regions includingThailand and causes a major problem to livestock productions. The majorsurface protein 5 (MSP5) is one of outer membrane protein of A. marginalewhich as an immunodominant protein encoded by a single gene and alsohighly conserved gene.Objective : The aim of this study was to optimize the conditions for the expression ofrecombinant 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. Therecombinant proteins were expressed by the plasmids in Escherichia colihost strain BL21 star™(DE3) at different temperatures (16, 25 and 37 C)for 6 h. The proteins were analyzed by SDS-PAGE and confirmed the targetprotein by Western blotting using antisera against His.Results : After induction with 0.1 mM of Isopropyl β-D-1-thiogalactopyranoside(IPTG) at different temperatures for protein expression, the protein wasnot produced at 6 h for 16 C. On the other hand, the rMSP5 protein wasproduced at 25 and 37 C for 2-6 h but the expressive protein at 25 Cshowed lower yield than that at 37 C.Conclusion : In this study, the best condition for rMSP5 protein expression wascultured at 37 C for 4 h. The protein was identified as the rMSP5 atthe molecular weight of 26 kDa.

DOI

10.58837/CHULA.CMJ.62.3.13

First Page

565

Last Page

573

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.