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Prokaryotic expression and characterisation of recombinant M1 protein of an Indian H5N1 avian influenza virus

Indian Agricultural Research Journals

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Title Prokaryotic expression and characterisation of recombinant M1 protein of an Indian H5N1 avian influenza virus
 
Creator SYED, ZOHRA
VENKATESH, GOVINDARAJULU
MURUGKAR, HARSHAD V
BEDEKAR, MEGHA KADAM
NAGARAJAN, SHANMUGASUNDARAM
DUBEY, SHIV CHANDRA
 
Subject Avian influenza, Characterization, Expression, Purification, Recombinant M1 protein
 
Description The type specific Matrix 1 (M1) protein of an Indian H5N1 avian influenza virus (AIV) was expressed as a histidine- tagged 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.
 
Publisher Indian Council of Agricultural Research
 
Date 2012-12-13
 
Type info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
 
Format application/pdf
 
Identifier https://epubs.icar.org.in/index.php/IJAnS/article/view/25653
10.56093/ijans.v82i12.25653
 
Source The Indian Journal of Animal Sciences; Vol. 82 No. 12 (2012); 1468–1471
2394-3327
0367-8318
 
Language eng
 
Relation https://epubs.icar.org.in/index.php/IJAnS/article/view/25653/11860
 
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