Expression of M Protein from LP02/C Equine Arteritis Virus Inhibits Growth of Escherichia Coli M15-pQE30 System

نویسندگان

  • Germán Ernesto Metz
  • María Mercedes Abeyá
  • María Soledad Serena
  • Carlos Javier Panei
  • Silvina Díaz
  • María Gabriela Echeverría
چکیده

Our objective was to obtain Equine Arteritis Virus M protein in prokaryotic system to test it as immunogen. LP02/C Equine Arteritis Virus cDNA was used as template to obtain and clone this protein. Equine Arteritis M protein was cloned in the expression vector pQE30 and the recombinant plasmid pQE30/M was transformed in Escherichia Coli M15 cells. The OD600 values of the IPTG-induced M15-pQE30/M culture showed an inhibition of the kinetics growth compared with the non-induced M15-pQE30/M and positive M15-pQE40/DHFR cultures. Several factors such as growth temperature, IPTG concentration and different inductors were analyzed but any of them showed an improvement in protein expression. Instead of E. coli M15strain, a new strain (E. coli BL21) was used and transformed with the pQE30/M. This resolved in part the growth inhibition observed in E. coli M15 cells, but no the recovery yield of the protein. So, as all gene products that affect cells kinetics growth are considered to be toxic, we argue that the lower yields in M protein recovery could be attributed to an associated toxicity of EAV-M protein from LP02/C strain in this expression system. Correspondence to: Dr. Germán Ernesto Metz, Assistant Researcher from CONICET-Argentina Virology, Faculty of Veterinary Sciences, National University of La Plata, La Plata, Argentina, Tel: +54 0221 4824956; Fax: +54 0221 4824956; E-Mail: [email protected]

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تاریخ انتشار 2017