Analisis Molekuler Gen Polymerase Basic 2 (PB 2) Virus Avian Influenza H5N1 yang Diisolasi dari Unggas Asal Purworejo, Jawa Tengah dan Bantul, Yogyakarta

Abstract

Avian Influenza Virus (AIV) is a RNA virus which can infect avian, mammalian, and human with genome consisting of 8 segments with Polymerase Basic 2 (PB2) as the longest segment. Polymerase Basic 2 has been known to play a role in cleavage and capping of RNA, related with replication and transcription into 8 segments of virus genome. A research with reverse genetic shows that Glutamate mutation into Lysine in position 627 PB2, determines the high pathogen and mutation in amino acid 667 into Lysine has been known as causing favorable virus to replicate at temperature of 360C. The goal of this study was to find out nucleotide of PB2 gene AIV subtype H5NI isolated from chicken Purworejo, Central Java and duck from Bantul, Yogyakarta. This study was also aimed to find out the existence of amino acid change in positions 627 and 667, homological level and its 3 dimension of protein structure. RNA virus isolated from the two test isolate was carried out RT-PCR (Reverse Transcriptase-Polymerase Chain Reaction) and the resulted cDNA which than sequencing to identify the possibility of the happening of change (mutation) in nucleotide sequence and its amino acid. The analysis was carried out by comparing a certain part of the sequence of nucleotide of PB2 protein of two test isolate with other H5N1 virus from avian and human coming from Asia, Europe, and Africa within 1996 – 2007 selected from genebank. Nucleotide sequence obtained was then analyzed filogenetic to find out its homological level, and subsequently carried out observation on its 3 dimension structure. The result of the study showed that amino acid in position of 627 and 667 in th two test isolate had not mutated into lysine. The mutation point was met in the residue of amino acid having fewer roles in replication, transcription, and transmission among species. Phylogenetic analysis had also been carried out by comparing the two test isolate of 62 isolates of avian influenza virus H5N1 from human and other avian. The two test isolate still showed high homological level with isolate from Indonesia, especially Yogyakarta. From the observation of 3 dimension structure, as a whole, the molecular characteristic of the two test isolate still showed the characteristic of amino acid residue generally found in avian from Indonesia and several avian isolates from other countries.