cloning & the sequencing of HAO3 gene ( Hyalomma Anatolicum Anatolicum) in secretory shuttle vector pPICZαA , Rigi cheshmehali  Garshasb , Aghayipor Khosrow , Mashayekhpour Saeid , Ebrahimi Seyed mahmoud , Razi Vaccine and Serum Research Institute

Abstract

     Hyalomma anatolicum anatulicum (Acari: Ixodidae) parasitize farm and wild animals in tropical and sub tropical region of the world. Ticks of the genus Hyalomma are well – khown vectors of viruses and avid parasits of man. One of the most important disease transmited by Hyalomma ticks is Crimean – Congo Hemorragic Fever (CCHF). H.anatolicum is one of the main vector ticks. In order to develop a broad – spectrum protection agianst different Hyalomma variants, some recent studies have been aimed at the HAO3 gene of Hyalomma anatolicum anatulicum.In this study, open reading frame of Hyalomma HAO3 gene was amplified by PCR using specific primers with restriction sites for XbaI and EcoRI restriction enzymes, cloned to pPICZαA expression vector in E.Coli DH5α. recimbinant vector was confirmed by PCR and restriction enzymes. Then cloned products were sent for sequencing to check the correct ORF and direct origin of the cloned HAO3 gene. Comparison of the nucleotid sequence of this gene and sequence of amino acid product of that with registered sequences in genbank shows that this gene with total isolated types HAO3, BM86, HA98 has much resemblance and after gene expression in eukaryotic yeast (pichia pastoris), it’s protein product can be a good candidate for recombinant vaccine against the Hyalomma.

 

Keywords: cloning-sequencing- PCR -HAO3gene- pPICZαA vector