Skip to main content
PMC home page
Springer Nature - PMC COVID-19 Collection logoLink to Springer Nature - PMC COVID-19 Collection
. 2005 Jan 1;30(1):69–73. doi: 10.1007/s11262-004-4583-z

Cloning and Sequence Analysis of the M gene of Porcine Epidemic Diarrhea Virus LJB/03

Fan Jinghui 1, Li Yijing 1,✉
PMCID: PMC7089338  PMID: 15744564

Abstract

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.

Keywords: cloning, M gene, nucleotide sequence, PEDV

References

  1. Liangxian Wu. Chinese FuJian J Anim Vet Sci. 2000;22(5):7. [Google Scholar]
  2. Weizhong Wu, Chinese Qinghai J. Anim Vet Sci. 2001;31(4):43–44. [Google Scholar]
  3. Yanxiu Ni, Jihuang Lin, Kongwang He, Honghua Huan. Anim Husbandry Vet Med. 2001;33(1):38–40. [Google Scholar]
  4. Barbara E. Straw., Sylvie D’llaire, William L. Mengeling, and David J. Taylor. Disease of Swine. 8th edn. Translated by Zhao Deming, Zhang Zhongqiu, Shen Jianzhong. 2000, pp. 181–187.
  5. Timoney J.F., Gillespie J.H., Scott F.W., Barlough J.E. Hagan and Bruner’s Microbiology and Infectious Diease of Domestic Animals . Ithaca: Comstock Publishing Associates; 1988. pp. 897–898. [Google Scholar]
  6. Bridgen A., Duarte M., Tobler K., Laude H., Ackermann M. J Gen Husbandry Virol. 1993;74:1798–1804. doi: 10.1099/0022-1317-74-9-1795. [DOI] [PubMed] [Google Scholar]
  7. Pensaert M.B., Straw B.E., D’Aallaire S., Mengeling W.L., Taylor D.I. Diease of Swine. Ames, IA: The Lowa University Press; 1999. pp. 179–185. [Google Scholar]
  8. Oldham, J., Pig Farming (Oct Suppl), 1972, pp. 72–73.
  9. DeBouck P., Callebaut P., Pensaert M. Proc Int Congr Pig Vet Soc. 1982;7:53. doi: 10.1016/0378-1135(82)90009-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hofmann M., Wyler R. Schweiz Arch Tierheilkd. 1987;129:437–442. [PubMed] [Google Scholar]
  11. Mostl K., Horvath E., Burki F. Wien Tierarztl Monatsschr. 1990; 77:10–18. [Google Scholar]
  12. Cheng Qinghua, Xiaoying Niu, Chengyu Ye. Chinese Qinghai J Anim Vet Sci. 1992; 22(3):22–23. [Google Scholar]
  13. Callebaut P. and DeBouck P., in Proc 5th. Int Congr Virol, Strasbonrg, p. 420.
  14. DeBouck P., Pensaert M. Am J Vet Res. 1980;41:219–223. [PubMed] [Google Scholar]
  15. Pijpers A., Van Nieuwstadt A.P., Terpstra C., Verheijden J.H.M. Vet Rec. 1993;132:129–131. doi: 10.1136/vr.132.6.129. [DOI] [PubMed] [Google Scholar]
  16. Wood E.N. Vet Res. 1977;100:243–244. doi: 10.1136/vr.100.12.243. [DOI] [PubMed] [Google Scholar]
  17. Murphy F.A., Gibbs E.P.J., HorZinek M.C., Studdert M.J., editors. Veterinary Virology. San Diego: Academic Press; 1999. pp. 496–501. [Google Scholar]
  18. Egberink H.F., Ederveen J., Callebant P., Horzinek M.C., Am J. Vet Res. 1988;49:1320–1324. [PubMed] [Google Scholar]
  19. Knuchel M., Ackermann M., Muller H.K., Kihm U. Vet Microbiol. 1992;32:117–134. doi: 10.1016/0378-1135(92)90100-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sang-Geon Yeo, Hernandez Mercedes, Krell Peter J. , Nagy Eva . Virus Genes. 2003; 26:239–246. doi: 10.1023/A:1024443112717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sambrook, J., David W., and Russell, Molecular Cloning: A Laboratory Manual, 3rd edn. Translated by Huang Peitang. 2002, pp. 27–30.

Articles from Virus Genes are provided here courtesy of Nature Publishing Group

RESOURCES