Sequencing of the F gene of Razi Institute's vaccinal NDV strain Clone IR 12

Document Type : Full Research Paper

Authors

1 Department of Poultry Research and Vaccine Production

2 Department of Avian Disease Research and Diagnostic, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Alborz 3197619751, Iran.

3 Department of Avian Disease Research and Diagnostic, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Alborz 3197619751, Iran

Abstract

Newcastle disease virus is an important virus in poultry industry. Use of killed or attenuated vaccine against the disease elevates immunological protection. The F gene is a major determinant in virulence of the virus. Therefore, in this study we aimed to sequence the F gene of Razi LaSota-derived and plaque-purified NDV strain IR12 and analyze it. The virus was inoculated into 10-day-old SPF embryonated chicken eggs and then the allantoic fluid was subjected to sucrose gradient purification using ultra centrifuge. The purified virus band was then subjected to RNA extraction and cDNA synthesis using gene specific primers. PCR was run using specific primers to amplify the F gene and then the band was gel extracted and cloned to pJet1.2 plasmids in a 3 to 1 ratio and transformed to a suitable competent cell. The extracted plasmid was then digested with BglII restriction enzyme to confirm the presence of the insert and then the plasmid was Sanger se As a result, the gene sequence F was completely sequenced and compared with the lentogenic strains found in the gene bank.quenced. The sequence was then assembled using MEGA6, the cleavage-site sequence of
112G-R-Q-G-R-L117 was obtained and then the phylogenetic tree was drawn. As a result, the gene sequence F was completely sequenced and compared with the lentogenic strains found in the gene bank.

Keywords

Main Subjects


1. Aldous, E.W. and D.J. Alexander .2001. Detection and differentiation of Newcastle disease virus (avian paramyxovirus type 1). Avian Pathology 30(2): p. 117-28.
2. Alexander, D.J., E.W. Aldous, and C.M. Fuller .2012. The long view: a selective review of 40 years of Newcastle disease research. Avian Pathology 41(4): p. 329-335.
3.Angel E. Absalón; Andrea Mariano; MatíasAlejandra; Vásquez-MárquezAndrés; Morales; Garzón Diana V. Cortés-Espinosa .2012. Complete genome sequence of a velogenic Newcastle disease virus isolated in Mexico. Virus Gene 45(2): p. 304-10.
4. de Leeuw, O.and Peeters .1999. Complete nucleotide sequence of Newcastle disease virus: for the existence of a new genus within the subfamily Paramyxovirinae, J. Gen. Virol 80 (Pt 1), 131-136.
5. Diego G. Diel, Leonardo Susta, Stivalis Cardenas Garcia, Mary L. Killian, Corrie C. Brown, Patti J. Miller, Claudio L. Afonso. 2012. Complete Genome and Clinicopathological Characterization of a Virulent Newcastle Disease Virus Isolate from South America. Journal of Clinical Microbiology 50(2): p. 378-387.
6. Ebrahimi MM, Shahsavandi S, Famil-Ghadakchi H, Masoudi S, Ghodsian N, Ebrahimi SR, Abdoshah M.2014. Clone Purification, Characterization and Standardization of LaSota Strain for Developing a Live Vaccine against Newcastle Disease Virus. Iranian Journal of Virology 8(4): p. 12-19.
7. Ganar K, Das M, Sinha S, Kumar S .2014. Newcastle disease virus: current status and our understanding. Virus Research 184:71–81.
8.Guoyuan Wen, Yu Shang, Jing Guo, Chen Chen, Huabin Shao, Qingping Luo .2013. Complete genome sequence and molecular characterization of thermostable Newcastle disease virus strain TS09-C. Virus Genes 46(3): p. 542-54.
9. Kavitha Murulitharan, Khadijah Yusoff, Abdul Rahman Omar, Aidin Molouki .2013. characterization of Malaysian velogenic NDV strain AF2240-I genomic sequence: a comparative study. Virus Genes 46(3): p. 431-40.
10. Bruschke C, Vallat B. 2008. OIE standards and guidelines related to trade and poultry diseases. Revue scientifique et technique. 27(3):62
11. Yurchenko KS, Sobolev IA, Glushchenko AV, Shestopalov AM. .2015. Complete Genome Sequence of Genotype Ib Newcastle Disease Virus Isolated from a Mallard (Anas platyrhynchos) in Russia. Genome Announc 3(6) pii: e01414-15.
12. Yurchenko KS, Zhou P, Kovner AV, Zavjalov EL, Shestopalova LV, Shestopalov AM. 2018. Oncolytic effect of wild-type Newcastle disease virus isolates in cancer cell lines in vitro and in vivo on xenograft model. PLoS ONE 13(4): e0195425.
13. Westbury H .2001. Newcastle disease virus: an evolving pathogen. Avian Pathology 30(1):5-11.