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In vitro expression of the recombinant fusion protein of Newcastle disease virus from local Indonesian isolates by using a cell-free protein expression system

Indonesian Journal of Biotechnology · Vol. 25 No. 2 (2020) · pp. 69–75

DOI
10.22146/ijbiotech.54703
ISSN
2089-2241, 0853-8654
Published
2020-11-10
Publisher
Universitas Gadjah Mada
Language
eng
License
creativecommons.org/licenses/by-sa/4.0

Abstract

The aim of this work was the in vitro expression of the recombinant fusion (F) protein of Newcastle disease virus (NDV). The pBT7-N-His-Fusion-NDV expression plasmid which carries the recombinant F protein encoding gene from local Indonesian isolates, was prepared and transformed into E. coli BL21 (DE3). To detect bacterial colonies carrying the recombinant plasmid, a restriction endonuclease analysis was performed using the EcoRI restriction endonuclease. These results showed that the pBT-N-His-Fusion-NDV plasmid was successfully isolated with a size of 4.601 bp, and three recombinant plasmids carrying the gene coding for the recombinant F protein of NDV were obtained. Selected recombinant plasmids were then in vitro by using a cell-free protein expression system followed by visualization of the recombinant F protein on a 12% SDS-PAGE gel both by Coomassie Brilliant Blue staining and Western blotting. Recombinant F protein was successfully in vitro expressed by using a cell-free protein expression system as indicated by a specific single protein band with a molecular mass of 25.6 kDa.

Keywords

Collaboration network of the authors

40 authors · 133 co-authorship links · showing the 40 most connected

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  • Universitas Gadjah Mada
  • Affiliation unknown
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Co-authorOrganisationJoint articles
Michael KannAffiliation unknown7
Tri WibawaUniversitas Gadjah Mada4
Krisna Noli AndrianAffiliation unknown4
Warih Pulung NugrahaniAffiliation unknown3
Ulayatul KustiatiAffiliation unknown3
Diana SavitriAffiliation unknown2
Irhamna PutriAffiliation unknown2
Medania PurwaningrumAffiliation unknown2
Teguh BudipitojoAffiliation unknown2
Dwi Liliek KusindartaAffiliation unknown2
Hera NirwatiAffiliation unknown2
S. SutaryoAffiliation unknown2
Nenny Sri MulyaniAffiliation unknown2
Titis WidowatiAffiliation unknown2
Purnomo HadiAffiliation unknown2
Devita Uzlah SibaraniAffiliation unknown1
Sherly GusnawanAffiliation unknown1
Lutfi MuzaqiAffiliation unknown1
Dyah SukendraAffiliation unknown1
Nungki Hapsari SuryaningtyasAffiliation unknown1
Beni ErnawanAffiliation unknown1
Wan Fatma ZuharahAffiliation unknown1
Yunita Dyah Puspita SantikAffiliation unknown1
Intan Tasya FarasyaAffiliation unknown1
Najiha RamadhaniAffiliation unknown1
Eldasari Myana Yulianingrum YulianingrumAffiliation unknown1
Rinawati RinawatiAffiliation unknown1
E SuharyantoAffiliation unknown1
Muhammad AdnanAffiliation unknown1
Matin NuhamunadaAffiliation unknown1
Lisna HidayatiAffiliation unknown1
Abdul Rahman SiregarAffiliation unknown1
Shinta HartantoAffiliation unknown1
Sofia MubarikaAffiliation unknown1
Camelia HerdiniAffiliation unknown1
Bambang HariwiyantoAffiliation unknown1
Akira HosoyamaAffiliation unknown1

Affiliations come from OpenAlex; journal OAI-PMH metadata carries none. Authors without an OpenAlex ID are matched by name, which can merge or split people.

Metadata source: OAI-PMH (metadata jurnal) https://jurnal.ugm.ac.id/ijbiotech/oai, first harvested 8 Nov 2026, last seen 8 Nov 2026.
Original metadata (oai_dc) 
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	<dc:title xml:lang="en-US">In vitro expression of the recombinant fusion protein of Newcastle disease virus from local Indonesian isolates by using a cell-free protein expression system</dc:title>
	<dc:creator>Haryanto, Aris</dc:creator>
	<dc:creator>Wihadmadyatami, Hevi</dc:creator>
	<dc:creator>Wijayanti, Nastiti</dc:creator>
	<dc:subject xml:lang="en-US">cell-free protein expression; in vitro expression; Newcastle diseases virus (NDV); recombinant F protein</dc:subject>
	<dc:description xml:lang="en-US">The aim of this work was the in vitro expression of the recombinant fusion (F) protein of Newcastle disease virus (NDV).  The pBT7-N-His-Fusion-NDV expression plasmid which carries the recombinant F protein encoding gene from local Indonesian isolates, was prepared and transformed into E. coli BL21 (DE3). To detect bacterial colonies carrying the recombinant plasmid, a restriction endonuclease analysis was performed using the EcoRI restriction endonuclease. These results showed that the pBT-N-His-Fusion-NDV plasmid was successfully isolated with a size of 4.601 bp, and three recombinant plasmids carrying the gene coding for the recombinant F protein of NDV were obtained. Selected recombinant plasmids were then in vitro by using a cell-free protein expression system followed by visualization of the recombinant F protein on a 12% SDS-PAGE gel both by Coomassie Brilliant Blue staining and Western blotting. Recombinant F protein was successfully in vitro expressed by using a cell-free protein expression system as indicated by a specific single protein band with a molecular mass of 25.6 kDa.</dc:description>
	<dc:publisher xml:lang="en-US">Universitas Gadjah Mada</dc:publisher>
	<dc:contributor xml:lang="en-US">DRPM Kemenristekdikti</dc:contributor>
	<dc:date>2020-11-10</dc:date>
	<dc:type>info:eu-repo/semantics/article</dc:type>
	<dc:type>info:eu-repo/semantics/publishedVersion</dc:type>
	<dc:type xml:lang="en-US">Original Research Articles</dc:type>
	<dc:format>application/pdf</dc:format>
	<dc:identifier>https://journal.ugm.ac.id/ijbiotech/article/view/54703</dc:identifier>
	<dc:identifier>10.22146/ijbiotech.54703</dc:identifier>
	<dc:source xml:lang="en-US">Indonesian Journal of Biotechnology; Vol 25, No 2 (2020); 69-75</dc:source>
	<dc:source>2089-2241</dc:source>
	<dc:source>0853-8654</dc:source>
	<dc:language>eng</dc:language>
	<dc:relation>https://journal.ugm.ac.id/ijbiotech/article/view/54703/29742</dc:relation>
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	<dc:relation>https://journal.ugm.ac.id/ijbiotech/article/downloadSuppFile/54703/13185</dc:relation>
	<dc:relation>https://journal.ugm.ac.id/ijbiotech/article/downloadSuppFile/54703/13186</dc:relation>
	<dc:relation>https://journal.ugm.ac.id/ijbiotech/article/downloadSuppFile/54703/13187</dc:relation>
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	<dc:rights xml:lang="en-US">Copyright (c) 2020 The Author(s)</dc:rights>
	<dc:rights xml:lang="en-US">http://creativecommons.org/licenses/by-sa/4.0</dc:rights>
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