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Detection of bone morphogenetic protein 2 within avian genome using PCR-based method

Bone Morhogenetic Protein 2 was first discovered by Urist (1965) when he implanted the demineralized bone in rat muscle pouch which result in bone formation at the implantation site. The presence of BMP 2 protein was further confirmed by Sampath and Reddi (1981) when they discovered the removal of B...

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Main Authors: Teh, Richard, Abu, Jalila, Omar, Abdul Rahman, Kumaran, J.
Format: Conference or Workshop Item
Language:English
Published: Faculty of Veterinary Medicine, Universiti Putra Malaysia 2013
Online Access:http://psasir.upm.edu.my/id/eprint/41435/1/41435.pdf
http://psasir.upm.edu.my/id/eprint/41435/
http://www.vet.upm.edu.my/dokumen/90301_proceeding_WPSA_V2_first_second_XX_new_20121013_%281%29.pdf
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spelling my.upm.eprints.414352016-01-28T02:19:53Z http://psasir.upm.edu.my/id/eprint/41435/ Detection of bone morphogenetic protein 2 within avian genome using PCR-based method Teh, Richard Abu, Jalila Omar, Abdul Rahman Kumaran, J. Bone Morhogenetic Protein 2 was first discovered by Urist (1965) when he implanted the demineralized bone in rat muscle pouch which result in bone formation at the implantation site. The presence of BMP 2 protein was further confirmed by Sampath and Reddi (1981) when they discovered the removal of BMP 2 protein caused the matrix failed to induce new bone formation. On the other hand, new bone formation was observed with the presence of BMP 2. Ever since then, the effects of BMP 2 has been actively study and lots of clinical trial have been done using recombinant BMP 2 protein produce from mammalian host but not from avian. Thus, the aim of this study is to detect the presence of BMP 2 gene within the avian genome. In order to amplify the BMP2 gene from avian genome, a set of primer was design based on the Gallus gallus nucleotides sequnces from NCBI (Accession: NM_204358.1). The two primers designed for this study were; forward primer (BMP2F) 5’-ATGGTTGCCGCCACCCGCTC-3’ and reverse primer (BMP2R) 5’-TCAGCGGCACCCGCAGCCCT-3’. DNA from few types of avian species were extracted from the respectively species feathers followed Richard and Jayaraj (2010) protocol and further subjected to PCR screening. The annealing temperature for this particular set of primer was set at 71.9°C. The PCR results were then send for sequencing. As shown in Figure 1.0, the PCR results in 1% agarose gel (w/v), the expected PCR products size was 1179 bps but the PCR product only shown about 950 bps instead of 1179 bps. However the sequencing results of these PCR products shown up to 94% similarity, when blast against the BMP 2 gene from NCBI. Despite the PCR products size were lesser than the expected size but it should be able to function in avian system as clinical report from Sample et al. (2008) had proven that recombinant human BMP 2 in calcium phosphate matrix as a bone graft substitute can be used for treatment of a comminuted, open humeral fracture in a whooping crane. Whereby, the Homo sapiens BMP 2 gene sequence (Accession: NM_001200.2) only shown 79% of similarity when blast against the Gallus gallus BMP 2 gene sequence. Faculty of Veterinary Medicine, Universiti Putra Malaysia 2013 Conference or Workshop Item PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/41435/1/41435.pdf Teh, Richard and Abu, Jalila and Omar, Abdul Rahman and Kumaran, J. (2013) Detection of bone morphogenetic protein 2 within avian genome using PCR-based method. In: World’s Poultry Science Association (Malaysia Branch) and World Veterinary Poultry Association (Malaysia Branch) Scientific Conference 2013, 30 Nov.-1 Dec. 2013, Faculty of Veterinary Medicine, Universiti Putra Malaysia. (pp. 142-143). http://www.vet.upm.edu.my/dokumen/90301_proceeding_WPSA_V2_first_second_XX_new_20121013_%281%29.pdf
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description Bone Morhogenetic Protein 2 was first discovered by Urist (1965) when he implanted the demineralized bone in rat muscle pouch which result in bone formation at the implantation site. The presence of BMP 2 protein was further confirmed by Sampath and Reddi (1981) when they discovered the removal of BMP 2 protein caused the matrix failed to induce new bone formation. On the other hand, new bone formation was observed with the presence of BMP 2. Ever since then, the effects of BMP 2 has been actively study and lots of clinical trial have been done using recombinant BMP 2 protein produce from mammalian host but not from avian. Thus, the aim of this study is to detect the presence of BMP 2 gene within the avian genome. In order to amplify the BMP2 gene from avian genome, a set of primer was design based on the Gallus gallus nucleotides sequnces from NCBI (Accession: NM_204358.1). The two primers designed for this study were; forward primer (BMP2F) 5’-ATGGTTGCCGCCACCCGCTC-3’ and reverse primer (BMP2R) 5’-TCAGCGGCACCCGCAGCCCT-3’. DNA from few types of avian species were extracted from the respectively species feathers followed Richard and Jayaraj (2010) protocol and further subjected to PCR screening. The annealing temperature for this particular set of primer was set at 71.9°C. The PCR results were then send for sequencing. As shown in Figure 1.0, the PCR results in 1% agarose gel (w/v), the expected PCR products size was 1179 bps but the PCR product only shown about 950 bps instead of 1179 bps. However the sequencing results of these PCR products shown up to 94% similarity, when blast against the BMP 2 gene from NCBI. Despite the PCR products size were lesser than the expected size but it should be able to function in avian system as clinical report from Sample et al. (2008) had proven that recombinant human BMP 2 in calcium phosphate matrix as a bone graft substitute can be used for treatment of a comminuted, open humeral fracture in a whooping crane. Whereby, the Homo sapiens BMP 2 gene sequence (Accession: NM_001200.2) only shown 79% of similarity when blast against the Gallus gallus BMP 2 gene sequence.
format Conference or Workshop Item
author Teh, Richard
Abu, Jalila
Omar, Abdul Rahman
Kumaran, J.
spellingShingle Teh, Richard
Abu, Jalila
Omar, Abdul Rahman
Kumaran, J.
Detection of bone morphogenetic protein 2 within avian genome using PCR-based method
author_facet Teh, Richard
Abu, Jalila
Omar, Abdul Rahman
Kumaran, J.
author_sort Teh, Richard
title Detection of bone morphogenetic protein 2 within avian genome using PCR-based method
title_short Detection of bone morphogenetic protein 2 within avian genome using PCR-based method
title_full Detection of bone morphogenetic protein 2 within avian genome using PCR-based method
title_fullStr Detection of bone morphogenetic protein 2 within avian genome using PCR-based method
title_full_unstemmed Detection of bone morphogenetic protein 2 within avian genome using PCR-based method
title_sort detection of bone morphogenetic protein 2 within avian genome using pcr-based method
publisher Faculty of Veterinary Medicine, Universiti Putra Malaysia
publishDate 2013
url http://psasir.upm.edu.my/id/eprint/41435/1/41435.pdf
http://psasir.upm.edu.my/id/eprint/41435/
http://www.vet.upm.edu.my/dokumen/90301_proceeding_WPSA_V2_first_second_XX_new_20121013_%281%29.pdf
_version_ 1643832996988452864
score 13.211869

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