Cloning And Sequence Analysis Of Complete Genome CDNA Of O Serotype Foot-and-Mouth Disease Virus SEA Genotype

Foot-and-Mouth Disease (FMD) is an extremely contagious disease inflicting cloven-hoofedanimals, which is caused by Foot-and-Mouth Disease Virus. Its occurrence and spreading seriouslyimpede animal husbandry development and foreign trade of related products. Therefore, muchattention has been paid to its study world-widely.Five fragments were amplified by RT-PCR according to FMDV genomic sequence andrestriction enzyme sites of clone vector. Complete genomic cDNA was generated by cloning,sequencing and assembly. T7 promoter sequence was added to of 5'leader untranslated region,specific restriction enzyme site was added to the 3' tailor untranslated region.Sequence alignmentwas carried out with sequences published, here reported, for the first time, the complete FMDVswine origin genomic sequence of serotype O SEA genotype (Strain HW04 ) was gene.Homologies were determined by comparison with sequences available in GenBank.Full-length genomic sequence was totally 8224bp, containing one ORF of 6999 nt, encoding apolyproteins composed of 2333 amino acids. Its initiation and stop codon were AUG and UAA,5'and 3'-UTR ( untranslated region) were 1095bp and 92bp,respectively. Gene L ,P1,P2 and P3were 603 bp,2208 bp,1464 bp and2721bp, respectively and 17-bp Poly (C), 38-bp Poly (A).VP1-based phylogenetie tree indicates: the homology between strain HW04 and strainO-MYA-7-98 was the highest, their nucleotide sequence and amino acid sequence homologies were93.3%,94.4%, respectively, so strain HW04 was confirmed belong to type O serotype SEA. Thehomology of L gene between strain HW04 and strain OMâ…¢-China was the highest, reaching to86,4%. 3A region of strain HW04 was 459 bp without any deletion, its amino acid had the highesthomology with O-HKN-17-82 of bovine origin.Potential protein decoration sites of strain HW04 at L and 3A were predicted using GeneRunner software: L protein has two glycosylation sites, so were the strain O-SKR-2002,O-TaiwanYunin,Tau-YuanTW97 of swine isolates, interestingly, these three strains were come from bovinetransmission. Through BLAST, 13 strains of swine isolates and 10 strains of bovine isolatessequences were available in GenBank,sequenee alignment reveals 10 strains of bovine isolates and4 strains of swine isolates (O-BUR-6-89,O-SKR-2002,O-CAM-1-98,and HW04) had completesequence without any deletion, while the other 10 strains of swine isolates had deletion spanningamino acid 92 to 101 in 3A.The above prediction suggests strain HW04 of swine isolate comesfrom bovinetransmission. The construction of full-length genomic cDNA clone of FMDV is the crucial step toobtain infectious clone, and the obtain of complete genomic cDNA provides substantial foundationfor further study of the structure and function of FMDV genome.