Processing of multimer FMD virus VP1-2A protein expressed in E. coli into monomers.
Expressions of several genes in bacteria were carried out by independent promoter. However, in case of eukaryotes ribosome skipping and introduction of IRES are employed as alternative to multiple translation initiation. Foot and mouth disease virus (FMDV) 2A peptide has been widely used for co-expression of multiple genes in eukaryotic, plant and mammalian systems. The 18 amino acid 2A peptide of FMDV facilitates efficient co-translational dissociation of the polyprotein into discrete protein products. To study the role of 2A in multimeric protein production a construct consisting of tandem repeat of 4 units of C- terminal VP1 linked through 2A sequence was made and expressed in E. coli. Along with tetramer protein, trimer, dimer and monomer proteins were produced. Stability studies showed that the tetramer protein was cleaved to smaller monomer on storage. The results provide scope for using FMDV 2A for expressing multiple genes under a single promoter in prokaryotes.
Dechamma
H J
HJ
FMD Research Lab, Indian Veterinary Research Institute, Hebbal, Bangalore 560 024, India. dechammahj@yahoo.com
Ashok Kumar
C
C
Nagarajan
G
G
Suryanarayana
V V S
VV
D000818:Animals; D003001:Cloning, Molecular; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D005536:Foot-and-Mouth Disease / Q000378:metabolism / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000378:metabolism*; D005786:Gene Expression Regulation; D015967:Gene Expression Regulation, Viral; D005821:Genetic Techniques; D010447:Peptide Hydrolases / Q000737:chemistry* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D011401:Promoter Regions, Genetic; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism
D010455:Peptides; D014764:Viral Proteins; D010447:Peptide Hydrolases