Cloning of a molecular vector encodes for the Hemagglutinin gene of the Egyptian strain of avian influenza A virus (H5N1) / Maye Mamdouh Abdulatif Khalifa ; Supervised Mohamed Akmal Elghor , Mohamed Mustafa Mustafa Ibrahim

Thesis (M.Sc.) - Cairo University - Faculty of Science - Department of Zoology

The spread of highly pathogenic avian H5N1 influenza viruses since 1997 and their virulence for poultry and humans has raised concerns about their potential to cause an influenza pandemic. Early detection of newly circulating viral strains as well as vaccination offers the most viable means to combat a pandemic threat. However, it will be a challenge to produce, distribute and implement a new vaccine if a pandemic spreads rapidly. The current study aimed at isolation of the full length hemagglutinin gene via Polymerase Chain Reaction (PCR), cloning the isolated gene and finally sequencing the amplified fragment in order to identify the restriction fragment considered for further expression cloning experiments. In order to achieve these objectives, a real time PCR protocol specific to the Egyptian H5N1 strains has been developed and tested on both in silico and in vitro levels. Moreover, bioinformatics analysis of generated DNA sequence data showed that the target H5N1 isolate is closely related to the other Egyptian H5N1 strains specially, strain (Egypt-MOH-NCR-7305). Finally, restriction recognition sequences inside the entire full length H5 gene have been identified

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