Abstract

Hrp pilus is a pathogenic bacterial surface appendage which serves as a tool for the transformation of bacterial effecter proteins into the plant cell. Hrp pilus is assembled by HrpA protein. In this study, a HrpA protein was successfully expressed, purified and a phage library single-chain fragment variable (scFv ) was constructed. After six rounds of bio-panning, a specific scFv against HrpA protein named scFv-FE5 was screened by phage display. The results of enzyme-linked immunosorbent assay (ELISA) and western blotting analysis showed that scFv-FE5 could recognize HrpA protein specifically. Current work demonstrates the preparation of the antibody anti-HrpA scFv-FE5 using phage display technology, a very useful tool for generation of a specific scFv to a certain antigen.

Key words: HrpA, expression, scFv, phage display, bio-panning.