Fig. 1 Flowchart of cloning, expression, and purification of recombinant protein. The gene of interest is cloned
into a cloning vector and subcloned into a bacterial expression vector. The recombinant constructs are used to
transform E. coli bacterial strains. Colonies containing the gene of interest are selected, and induction protein
expression is performed. After checking if the proteins are expressed in their soluble or insoluble fraction, a
large-scale production is completed for protein purification. Using a Ni
2+ -charged chromatography column,
the recombinant proteins are purified at their native or denatured condition. Purified samples are assessed by
SDS-PAGE
Cell Adhesion Assay for Leptospiral Proteins
179
into a cloning vector and subcloned into a bacterial expression vector. The recombinant constructs are used to
transform E. coli bacterial strains. Colonies containing the gene of interest are selected, and induction protein
expression is performed. After checking if the proteins are expressed in their soluble or insoluble fraction, a
large-scale production is completed for protein purification. Using a Ni
2+ -charged chromatography column,
the recombinant proteins are purified at their native or denatured condition. Purified samples are assessed by
SDS-PAGE
Cell Adhesion Assay for Leptospiral Proteins
179