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The calcium‐binding protein of Entamoeba histolytica as a fusion partner for expression of peptides in Escherichia coli
Author(s) -
Reddi Honey,
Bhattacharya Alok,
Kumar Vijay
Publication year - 2002
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20020024
Subject(s) - escherichia coli , fusion protein , entamoeba histolytica , biology , biochemistry , enteropeptidase , protein a/g , recombinant dna , microbiology and biotechnology , chemistry , gene
We describe the construction of an Escherichia coli expression vector, CBP that allows the C‐terminal fusion of heterologous proteins to the calcium‐binding protein (CaBP) of the parasitic protozoan Entamoeba histolytica. The intrinsic nature of this protein to remain soluble on heat treatment has been exploited in its use as a novel fusion partner. The presence of a histidine tag and an enterokinase recognition site, aid in the affinity purification and proteolytic cleavage of the fusion protein. The efficacy of the vector was tested using the preS1 region of the envelope protein of the hepatitis B virus. The CaBP‐preS1 fusion protein partitioned in the soluble fraction on heat treatment and this facilitated its rapid purification.