short communication: molecular characterization and...

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Pakistan J. Zool., vol. 49(x), pp xxx-xxx, 2017. DOI: http://dx.doi.org/10.17582/journal.pjz/2017.x.x.xxx.xxx * Corresponding author: [email protected] 0030-9923/2016/0004-1161 $ 8.00/0 Copyright 2016 Zoological Society of Pakistan Supplementary Table I.- Primer sequences used for identification and amplification of Fusion and Hemagglutinin gene along with their expected product sizes. No. Primer sequences (5ʹ - 3ʹ) Position on viral genome Product size (bp) Reference 1 Sense: GTGAAYTTTGTCTCCTTGAC Anti-sense: GAGGCATGTGCRAAAGC 3833-3852 to 4814- 4798 965 bp Munir et al., 2012 2 Sense: TTGAYGGCAGGCCTCTTG Anti-sense: GTGATAGAAGARCTTGACACCTC 4673-4690 to 5585-5563 890 bp 3 Sense: ATAATATGCGTGCCACCTA Anti-sense: ATAYACGGGTAGAACGGT 5465-5483 to 6370-6353 888 bp 4 Sense: TGGCTTGGGAAYAATACCCT Antisense: TGCAGTGTGAGTGCAACT 6190-6209 to 7176-7159 969 bp 5 Sense: GGGAGGCATACAACAGGACA Antisense: TGGTTGCAGCAATGCTCTC 289-308 to 512-530 242 bp Ling et al., 1997 Primer 1 to 4, used for amplification of F and HN gene of study isolate; Primer 5, used for the identification of NDV isolate. Supplementary Material Pakistan J. Zool., vol. 49(2), pp 755-759, 2017. DOI: http://dx.doi.org/10.17582/journal.pjz/2017.49.2.sc9 Short Communication: Molecular Characterization and Epitope Mapping of Fusion (F) and Hemagglutinin (HN) Genes of Avian Paramyxovirus Serotype I from Peacocks in Pakistan Sameera Akhtar 1 *, Muhammad Akram Muneer 1 , Khushi Muhammad 1 , Muhammad Yasin Tipu 1 , Muhammad Anees 2 , Imran Rashid 1 , Raza-ur- Rehman 3 and Irshad Hussain 1 1 University of Veterinary and Animal Sciences, Lahore 54000, Pakistan 2 Veterinary Research Institute, Lahore 54000, Pakistan 3 Poultry Research Institute, Rawalpindi 46000, Pakistan

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Page 1: Short Communication: Molecular Characterization and ...researcherslinks.com/uploads/currentIssues/SuppMaterial/1491609862Supp...2 Supplementary Fig. 1Phylogenetic consensus tree for

Pakistan J. Zool., vol. 49(x), pp xxx-xxx, 2017. DOI: http://dx.doi.org/10.17582/journal.pjz/2017.x.x.xxx.xxx

* Corresponding author: [email protected]/2016/0004-1161 $ 8.00/0Copyright 2016 Zoological Society of Pakistan

Supplementary Table I.- Primer sequences used for identification and amplification of Fusion and Hemagglutinin gene along with their expected product sizes.

No. Primer sequences (5ʹ - 3ʹ) Position on viral genome Product size (bp) Reference

1 Sense: GTGAAYTTTGTCTCCTTGACAnti-sense: GAGGCATGTGCRAAAGC

3833-3852 to 4814- 4798 965 bp Munir et al., 2012

2 Sense: TTGAYGGCAGGCCTCTTGAnti-sense: GTGATAGAAGARCTTGACACCTC

4673-4690 to 5585-5563 890 bp

3 Sense: ATAATATGCGTGCCACCTAAnti-sense: ATAYACGGGTAGAACGGT

5465-5483 to 6370-6353 888 bp

4 Sense: TGGCTTGGGAAYAATACCCTAntisense: TGCAGTGTGAGTGCAACT

6190-6209 to 7176-7159 969 bp

5 Sense: GGGAGGCATACAACAGGACAAntisense: TGGTTGCAGCAATGCTCTC

289-308 to 512-530 242 bp Ling et al., 1997

Primer 1 to 4, used for amplification of F and HN gene of study isolate; Primer 5, used for the identification of NDV isolate.

Supplementary Material

Pakistan J. Zool., vol. 49(2), pp 755-759, 2017. DOI: http://dx.doi.org/10.17582/journal.pjz/2017.49.2.sc9

Short Communication: Molecular Characterization and Epitope Mapping of Fusion (F) and Hemagglutinin (HN) Genes of Avian Paramyxovirus Serotype I from Peacocks in Pakistan

Sameera Akhtar1*, Muhammad Akram Muneer1, Khushi Muhammad1, Muhammad Yasin Tipu1, Muhammad Anees2, Imran Rashid1, Raza-ur-Rehman3 and Irshad Hussain1

1University of Veterinary and Animal Sciences, Lahore 54000, Pakistan2Veterinary Research Institute, Lahore 54000, Pakistan3Poultry Research Institute, Rawalpindi 46000, Pakistan

Page 2: Short Communication: Molecular Characterization and ...researcherslinks.com/uploads/currentIssues/SuppMaterial/1491609862Supp...2 Supplementary Fig. 1Phylogenetic consensus tree for

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Supplementary Fig. 1Phylogenetic consensus tree for the pigeon-originated NDV isolate for fusion gene (A), hemagglutinin gene (B) and hyper-variable region of F gene (C).The nucleotide sequences of study isolate for each gene were compared with corresponding genes of representative strains reported previously to public database, the GenBank. The evolutionary history was inferred using the Neighbor-Joining method with 1000 bootstrap value in MEGA version 6.0.

S. Akhtar et al.

Page 3: Short Communication: Molecular Characterization and ...researcherslinks.com/uploads/currentIssues/SuppMaterial/1491609862Supp...2 Supplementary Fig. 1Phylogenetic consensus tree for

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Supplementary Fig. 2.Alignment of deduced amino acid sequence of complete F gene of peacock isolate. The residue profile of study isolate is compared with strains of NDVs representing different genotypes including the vaccine strains. Structurally and functionally important residues are boxed and highlighted.

Molecular Characterization and Epitope Mapping of Fusion and Hemagglutinin 3

Page 4: Short Communication: Molecular Characterization and ...researcherslinks.com/uploads/currentIssues/SuppMaterial/1491609862Supp...2 Supplementary Fig. 1Phylogenetic consensus tree for

4 S. Akhtar et al.

Supplementary Fig. 3. Alignment of deduced amino acid sequence of complete HN gene of pigeon isolate. The residue profile of study isolate is compared with strains of NDVs representing different genotypes including the vaccine strains. Structurally and functionally important residues are boxed and highlighted.sequence of complete HN gene of pigeon isolate. The residue profile of study isolate is compared with strains of NDVs representing different genotypes including the vaccine strains. Structurally and functionally important residues are boxed and highlighted.