Comparative sequence analysis of genome segment 10 of African horsesickness virus type 4 to the cognate genes
Abstract: The genomic double stranded (ds) RNA of African horsesickness virus (AHSV) type 4SP was used as template to generate cDNA by random primed reverse-transcriptase reaction. Full-length copies of cDNA corresponding to the s10 gene of AHSV were amplified by polymerase chain reaction using primers to the conserved end sequences of the gene segment. S.10 cDNA was lighated into a TA cloning vector (PCR tm II) which was subsequently used to transform component Ecoli (NVaF). Proper ligation and transformation was assessed by growth of white colonies from X-GAL paltes, exctraction of plasmids with EcorR1. Agarose gel electrophoresis was used to ensure the size of the insert corresponded to full length S10 eDNA (720bp). Several positive clones were grown up to obtain an adequate amount of plasmid DNA for sequence analysis by PEG precipitation. The nucleotide of S10 cDNA was determined by dideoxy reaction using Sequenase version2.0 This study indicated that there is a considerable degree of homology between the S10 gene nucleotide sequence of AHSV type 4SP and the published sequence of the cognate gene. Furtheremore, while generally confiriming relationships between S10 sequences previously reported, the lack of correspondence between serotype and sequence relationships suggests that reassortment of this segment may have occurred in the AHSV 4SP isolate and indicates a potential problem in using serotype as classification variable for genes other than those coding for proteins directly involved in virus neutralization. The divergence of these sequence suggests a role for the S 10 gene in the differences in replication and pathogenesis seen between AHSV isolates.
Publication year 1997
Pages 986-1000
Availability location معهد بحوث الامصال واللقاحات البيطرية
Availability number
Organization Name
City cairo
serial title 23rd arabveterinary medical congress
Author(s) from ARC
Agris Categories Animal diseases
Proposed Agrovoc Horses-Diseases;
Publication Type Conference/Workshop
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