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.

کلیدواژه ها:

Mycoplasma agalactiae ، Expression ، Purification ، Recombinant protein ، Chimeric