The current outbreak of emerging microbial pathogens worldwide has stressed the importance and urgency of molecular surveillance of viruses. The rapid, specific and sensitive methods for detection of viruses playa key role in the success of control and prevention of virus infections. A number of methods have been developed to detect viruses, including immunological assays, transmission electron microscopy and PCR-based viral nucleic acid detection, and even optical microscopes. However, most of them are time consuming and require purifying the samples. Therefore, researchers are moving toward a potential approach which applies nanotechnology for development of biosensor to improve the sensitivity and detection of viruses. Herein, the authors report the results on expression of beta-subunit of the Bacillus SP3 F0F1-ATPase in E. coli and production of antibody against the recombinant protein. In order to achieve this, the DNA sequence encoding beta-subunit was amplified directly from genomic DNA of Bacillus SP3 by PCR, cloned into pCR2.1 vector and analyzed by restriction enzymes and DNA sequencing. This domain was then constructed into pET2la+ vector for expression in E. coli BL21 (DE3) strain. The recombinant beta-subunit protein was purified by ProBond™ NickelChelating Resin (Invitrogen) and was used to produce its polyclonal antibodies in rabbit. Five days after the last injection, rabbit was sacrified and the total blood was collected. The serum was separated from the total blood and stored at -80°C until use. The Western blot analysis showed that the antibodies reacted specifically toward beta-subunit of F0F1-ATPase present in chromatophore extracted from Rhodospirillum rubrum. These polyclonal antibodies have a potential to develop a biosensor to detect pathogens including viruses, bacterium and tumor cancer.
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