Sequence Analysis of the Haemagglutinin and Fusion Protein Genes of Peste-des-petits Ruminants Vaccine Virus of Indian Origin

Abstract

The amino acid composition of the two surface proteins of peste-des-petits ruminants vaccine virus belonging to lineage four from India were deduced from the nucleotide sequence. The fusion (F) protein gene of PPRV Sungri/96 is 2405 nucleotides long and in relation to the length, it is 80 nucleotides longer than that of PPRV Nigeria/75/1 which are found to be present at the 5'UTR of this virus. The complete F gene alignment with other morbillivirus reveals a homology of 89% with PPRV/Nigeria/75/1 and 48-51% with other morbilliviruses. The F protein of PPRV Sungri/96 exhibited characteristics similarity to those of other morbillivirus F proteins. The overall amino acid similarity with its counterpart PPRV Nigeria/75/1 was 96%; with other morbilliviruses it is 65-74%. The PPRV Sungri/96 haemagglutinin (H) protein gene is 1954 nucleotides long and showed a sequence homology of 90.7% with PPRV/Nigeria/75/1 and with other morbilliviruses it ranged from 33% to 45%. At amino acids level, PPRV Sungri/96 showed a homology of 92.3% with PPRV/Nigeria/75/1 and 34-49% with other morbilliviruses. The phylogenetic tree constructed for F and H gene reveals four separate groups which is very similar to that found in other genes. To the best of our knowledge this is the first report describing the F and H genes of an Indian isolate.

Development and Characterization of a Stable Vero Cell Line Constitutively ExpressingPeste des Petits Ruminants Virus(PPRV) Hemagglutinin Protein and Its Potential Use as Antigen in Enzyme-Linked Immunosorbent Assay for Serosurveillance of PPRV

“…3A, lane 1), with no such protein band in the normal Vero cell lysate (lane 2), indicating expression of PPRV H protein in stable cells. The calculated size of the PPRV H protein along with the His tag at the C-terminal end comes to around 69.98 kDa, which is in agreement with the mobility of the additional protein band observed in SDS-PAGE, indicating that the 70-kDa protein is the product from the cloned PPRV H gene (8).…”

“…This was achieved by employing the expression vector cassette of PPRV H gene under a CMV promoter-DNA construct with integration into the Vero cell genome. The sequence coding for PPRV H protein was amplified by RT-PCR, and the size of the amplified product (1,869 bp) and its nested PCR amplicon (718 bp) were in agreement with the reported sizes of the PPRV H gene (8,21). The amplicon cloned into the pTargeT vector was screened by colony PCR using T7 (Vector specific) and PPRV H (NotI) R His (virus specific) primers, which resulted in amplification of a 1,926-bp product, confirming the presence of an insert in the VOL.…”

Sequence and Phylogenetic Analyses of the Structural Genes of Virulent Isolates and Vaccine Strains of Peste Des Petits Ruminants Virus from India

“…Haemagglutinin protein of Indian origin of PPRV shares an identity of 89.6–98% and 89.8–97.9% at nt and aa levels with other PPRV, but with other morbilliviruses it ranges from 47.5–58.2% and 35–49% (Table 3). This indicates the high‐level divergence of the H protein among all the morbilliviruses (Tsukiyama et al., 1987; Dhar et al., 2006; Chard et al., 2008). The alignment (Fig.…”

“…Unique substitutions in PPRV isolates at positions E243D and S304A were noticed in comparison with other morbillivirus. In the non‐conserved domain (aa 485–517), which includes a hydrophobic anchor membrane sequence (aa 485–502), only two aa variations (in ICV 89 three aa) could be observed between the Asian and African lineages of PPRVs (Meyer and Diallo, 1995; Dhar et al., 2006). A total of 12 cysteine residues (except in ICV 89 isolate) are conserved across morbilliviruses, indicating the vital role of this aa in maintaining the tertiary structure (Barrett et al., 1993).…”

Peste des petits ruminants vaccine and vaccination in India: sharing experience with disease endemic countries

“…Entire genome of this Asian lineage vaccine virus has been sequenced [27]. Detailed comparative analysis of different genes of this vaccine virus is available in public domain [10,21,22]. Further, in order to increase the utility of PPR vaccine virus and to reduce the cost of vaccinations for the control of three important viral diseases of small ruminants, both sheep pox vaccine and goatpox vaccine have been used as combined vaccines with PPRV/sungri-96 Vaccine virus with encouraging results and field applications [8, ].…”