Highly Expression and Purification of Chimeric Protein P48-P40of Mycoplasma Agalactiae In E.Coli, Applicable For ELISA Test

Mycoplasma agalactiae is an economically important pathogen and the main etiological agent of contagious agalactia (CA) in small ruminants with a relatively high prevalence in sheep and goat in Iran, mainly characterized by mastitis, conjunctivitis and arthritis as predominant symptoms of a localized infection. Two genes encoding p48 and p40 major surface lipoprotein are played a fundamental role in the pathogenesis of Mycoplasma.ObjectivesIn this study a chimeric protein P48-p40 of Mycoplasma agalactiae was designed, expressed and purified which is applicable for some serological tests such as rELISA or a recombinant vaccine.Methods Complete coding sequence of p48-p40 wascloned into the expression vector Pet32a+ then transformed and expressed into E.coli BL 21 (DE3). The purification of TRX-p48-p40-6His was optimized by using Nickel Resin inaffinity batch formation method. ResultsAfter some experimental trials the designed chimeric protein was expressed successfully at optimal temperature 22oC after 4 hourswith 0.1mmolIPTG as an inducer. The protein was highly purified in soluble form by affinity batch formation method with yield of 25mg/L.Further study is under investigationto evaluate the function of chimeric protein in ELISA test, and also analysis of immune response in the animal model and the main host.

Highly Expression and Purification of Chimeric Protein P48-P40of Mycoplasma Agalactiae In E.Coli, Applicable For ELISA Test Keywords:

Mycoplasma agalactiae , Expression , Purification , Recombinant protein , Chimeric