Serological and Molecular Relationships of Two Egyptian Cyanophage Isolates

Abstract

This research aimed to investigate the serological relationship between two Egyptian cyanophage isolates (virulent and temperate) infecting Anabaena sp. A simple purification procedure, based on ultracentrifugation of the clarified lysates was applied, and diagnostic antise-ra were prepared by immunizing New-Zealand (NZ) rabbits using the purified preparations. Ouchterlony and micropreciptin tests were per-formed to check the serological relationship as well as discontinuous non-denaturing gel analysis. Molecular analysis of the amino acid se-quences of coat protein genes was applied using DNA star lasergene 10 suite. The purified preparations revealed A260/A280 ratios of 1.4 and 1.1, concentration was 1.005 and 0.58 mg/ml, and precipitation dilution end point of 1/32 and 1/16 for virulent and temperate isolate respectively. The purified suspensions (7x 104 and 5x 104 pfu/ml, estimated by plaque assay, for virulent and temperate isolate respectively) were used to obtain the specific antisera. The purified preparations are found immuno-genic, producing specific polyclonal antibodies. Data indicated a serological relationship between the two isolates which was discussed on the basis of discontinuous non-denaturing gel elec-trophoresis analysis and bioinformatic analysis of coat protein genes i.e antigenic indexes, secondary,3D and space fill structures.