Cloning and sequence analysis of the M gene of porcine epidemic diarrhea virus LJB/03.
Porcine epidemic diarrhea virus (PEDV) LJB/03 was isolated from the fece of piglets infected with PEDV on a pig farm, Heilongjiang province, China. The M gene of LJB/03 was amplified from the RNA extracted directly from the fece samples by RT-PCR and cloned into pMD18-T vector. The M gene cDNA was sequenced and encompasses an open reading frame of 681 nucleotides, encoding a 226-amino acid protein. The LJB/03 M gene has a base composition of 152 adenines (22%), 153 cytosines (23%), 161 guanines (24%), and 214 thymines (31%). Sequence comparison with other PEDV strains selected from GenBank revealed that the LJB/03 M gene has a high sequence homology to those of other PEDV isolates, 97.80% with JMe2, 96.92% with KPEDV-9 (Korean field isolate), 97.36% with KPEDV-9 (Korean), 97.80% with Br1/87, and 97.94% with CV777. The encoded protein shared 97.79% amino acid identities compared with CV777, 97.35% with Br1/87, 97.79% with JMe2, 96.90% with KPEDV-9 (Korean field isolate), 96.46% with KPEDV-9 (Korean). Sequence analysis of the M gene, including genetic distance measurement, phylogenetic tree analysis, and residue substitution analysis, showed that all other PED viruses analyzed fell into three groups, and the LJB/03 itself branched in an independent group. These data revealed that the M gene nucleotide sequence of LJB/03 has some mutations in comparison with the other PED viruses.
Jinghui
Fan
F
Veterinary Department, Northeast Agriculture University, Harbin, 150030, P. R. China.
Yijing
Li
L
D000818:Animals; D001482:Base Composition; D002681:China; D003001:Cloning, Molecular; D017934:Coronavirus / Q000235:genetics* / Q000302:isolation & purification; D018352:Coronavirus Infections / Q000662:veterinary / Q000821:virology; D000086862:Coronavirus M Proteins; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D005243:Feces / Q000821:virology; D005814:Genes, Viral; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D016366:Open Reading Frames / Q000235:genetics; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013552:Swine / Q000821:virology; D013553:Swine Diseases / Q000821:virology; D014763:Viral Matrix Proteins / Q000235:genetics*
D000086862:Coronavirus M Proteins; D018076:DNA, Complementary; C000706236:M protein, Porcine epidemic diarrhea virus; D012367:RNA, Viral; D014763:Viral Matrix Proteins
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