Prokaryotic expression and characterisation of recombinant M1 protein of an Indian H5N1 Avian Influenza virus

Abstract

The type specific Matrix 1 (M1) protein of an Indian H5N1 avian influenza virus (AIV) was expressed as a histidinetagged fusion protein in a prokaryotic expression system and characterized. The M1 gene was amplified by reverse transcription PCR using appropriately designed primers and cloned into the expression vector, pET28a ( + ). The orientation and reading frame of the recombinant expression construct (pET-M1) was confirmed by sequence analysis. The derived amino acid sequence homology between M1 of AIV H5N1 and other reference AIV subtypes was found to be 93.7% to 99.2%. The 33kDA recombinant M1 (rM1) protein was expressed as inclusion body after induction with 1 mM IPTG in E. coli BL21 (DE3)pLysS cells. The protein was purified to near homogeneity by affinity chromatography using Ni- NTA agarose column. The yield of the purified rM1 was found to be 2 mg/100 ml of induced culture. The rM1 was found to react specifically with H1-H15 AIV subtype specific sera in Western blot. The results indicated that the purified rM1 protein could be used as antigen for detection of type specific AIV antibodies by immunoassays.

Prokaryotic expression and characterisation of recombinant M1 protein of an Indian H5N1 avian influenza virus. Available from: https://www.researchgate.net/publication/283134347_Prokaryotic_expression_and_characterisation_of_recombinant_M1_protein_of_an_Indian_H5N1_avian_influenza_virus [accessed Jun 3, 2017].