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The Effect of Single Co-expression of The DnaK-DnaJ-GrpE and GroEL/ES Chaperones and Their Combination on Expression Intein-pretrombin-2 in Escherichia coli ER2566

VALENSI · Vol. ume No. 1 (2020) · hlm. 47–54

DOI
10.15408/jkv.v6i1.11333
ISSN
2548-3013, 2460-6065
Terbit
2020-05-29
Penerbit
Department of Chemistry, Faculty of Science and Technology Syarif Hidayatullah Jakarta State Islamic University
Bahasa
eng

Abstrak

The use of recombinant thrombin in the manufacture of fibrin glue allows diseases contamination to be avoided. However, the expression of recombinant protein in E. coli still has a disadvantage of the formation of inclusion bodies, so it needs to be minimized by co-expression of chaperones. Therefore, the aim of this study was to determine the effect of single DnaK-DnaJ-GrpE and GroEL/ES chaperone expression and their combination on the expression of intein-pretrombin-2Ti,pH on E. coli ER2566. The method started with isolation of pTWIN1-prethrombin-2Ti,pH and pG-KJE8 from E. coli TOP10F' and DH5α respectively, the co-transformation of the expression host E. coli ER2566 using pG-KJE8 and pTWIN1-prethrombin-2Ti,pHvectors, the chaperone co-expression was induced using L-Arabinosa before IPTG induction and cell culture growth was incubated at 22 oC. The expression products were characterized by using Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The results of the co-expression of chaperone showed that the number of soluble fraction was higher than the one without co-expression of chaperone. In addition, the co-expression of chaperone using pG-KJE8 in intein-prothrombin-2Ti,pH expression was sufficient using tetracycline as an inducer.

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  • Padjadjaran University
  • Universitas Jenderal Achmad Yani
  • National Research and Innovation Agency
  • Maranatha Christian University
  • Sekolah Tinggi Farmasi Indonesia
  • Universitas Bhakti Kencana
  • Sunan Gunung Djati State Islamic University Bandung
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Rekan penulisOrganisasiArtikel bersama
Muhammad YusufPadjadjaran University5
Shinta KusumawardaniPadjadjaran University2
Umi BarorohSekolah Tinggi Farmasi Indonesia2
Fauzian Giansyah RohmatullohPadjadjaran University2
Rahmaniar MulyaniPadjadjaran University2
Khomairi HasanUniversitas Jenderal Achmad Yani2
Mamlikatu Ilmi AzizahPadjadjaran University2
Wanda DestiaraniPadjadjaran University2
Ahmad Fariz MaulanaPadjadjaran University2
Rima HandiyaniPadjadjaran University1
Idar IdarUniversitas Bhakti Kencana1
Yani SuryaniSunan Gunung Djati State Islamic University Bandung1
Opik TaupiqurrohmanSekolah Tinggi Analis Bakti Asih1
Sukma NuswantaraOrganisasi Riset Hayati dan Lingkungan1
Ellsie Viendra PermanaUniversitas Jenderal Achmad Yani1
Yudith Yunia KusmalaUniversitas Jenderal Achmad Yani1
Iis Inayati RakhmatUniversitas Jenderal Achmad Yani1
Teja KoswaraUniversitas Jenderal Achmad Yani1
Mia Tria NoviantiPadjadjaran University1
Sayu Putu Yuni ParyatiUniversitas Jenderal Achmad Yani1
Retno PangastutiMaranatha Christian University1
Meilinah HidayatMaranatha Christian University1
Sijani PrahastutiMaranatha Christian University1
Maria UlfahNational Research and Innovation Agency1
Is HeliantiNational Research and Innovation Agency1
Niknik NurhayatiNational Research and Innovation Agency1
Gabriela Christy SabbathiniNational Research and Innovation Agency1
Sri Rezeki WulandariNational Research and Innovation Agency1
I Gede Eka Perdana PutraNational Research and Innovation Agency1
Hesti Lina WiraswatiPadjadjaran University1
Nisa FauziahPadjadjaran University1
Siti SoidahPadjadjaran University1
Sari SyahruniPadjadjaran University1
Henry ChandraPadjadjaran University1
Dhiya Ulhaq SalsabilaPadjadjaran University1
Bachti AlisjahbanaDr. Hasan Sadikin General Hospital1

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Sumber metadata: OAI-PMH (metadata jurnal) http://journal.uinjkt.ac.id/index.php/valensi/oai, pertama di-harvest 10 Nov 2026, terakhir terlihat 10 Nov 2026.
Metadata asli (oai_dc) 
<oai_dc:dc xmlns="http://www.openarchives.org/OAI/2.0/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:oai_dc="http://www.openarchives.org/OAI/2.0/oai_dc/" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xsi:schemaLocation="http://www.openarchives.org/OAI/2.0/oai_dc/  http://www.openarchives.org/OAI/2.0/oai_dc.xsd">
	<dc:title xml:lang="en">The Effect of Single Co-expression of The DnaK-DnaJ-GrpE and GroEL/ES Chaperones and Their Combination on Expression  Intein-pretrombin-2 in Escherichia coli ER2566</dc:title>
	<dc:creator xml:lang="en">Maksum, Iman Permana</dc:creator>
	<dc:creator xml:lang="en">Wildan, D Agus Yusuf</dc:creator>
	<dc:creator xml:lang="en">Hasan, Khomaini</dc:creator>
	<dc:creator xml:lang="en">Subroto, Toto</dc:creator>
	<dc:subject xml:lang="en">Intein-pretrombin-2Ti</dc:subject>
	<dc:subject xml:lang="en">pH</dc:subject>
	<dc:subject xml:lang="en">chaperone co-expression of DnaK-DnaJ-GrpE and GroEL/ES</dc:subject>
	<dc:subject xml:lang="en">E. coli ER2566</dc:subject>
	<dc:description xml:lang="en">The use of recombinant thrombin in the manufacture of fibrin glue allows diseases contamination to be avoided. However, the expression of recombinant protein in E. coli still has a disadvantage of the formation of inclusion bodies, so it needs to be minimized by co-expression of chaperones. Therefore, the aim of this study was to determine the effect of single DnaK-DnaJ-GrpE and GroEL/ES chaperone expression and their combination on the expression of intein-pretrombin-2Ti,pH on E. coli ER2566. The method started with isolation of pTWIN1-prethrombin-2Ti,pH and pG-KJE8 from E. coli TOP10F' and DH5α respectively, the co-transformation of the expression host E. coli ER2566 using pG-KJE8 and pTWIN1-prethrombin-2Ti,pHvectors, the chaperone co-expression was induced using L-Arabinosa before IPTG induction and cell culture growth was incubated at 22 oC. The expression products were characterized by using Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The results of the co-expression of chaperone showed that the number of soluble fraction was higher than the one without co-expression of chaperone. In addition, the co-expression of chaperone using pG-KJE8 in intein-prothrombin-2Ti,pH expression was sufficient using tetracycline as an inducer.</dc:description>
	<dc:publisher xml:lang="en">Department of Chemistry, Faculty of Science and Technology Syarif Hidayatullah Jakarta State Islamic University</dc:publisher>
	<dc:date>2020-05-29</dc:date>
	<dc:type>info:eu-repo/semantics/article</dc:type>
	<dc:type>info:eu-repo/semantics/publishedVersion</dc:type>
	<dc:format>application/pdf</dc:format>
	<dc:identifier>https://journal.uinjkt.ac.id/index.php/valensi/article/view/11333</dc:identifier>
	<dc:identifier>10.15408/jkv.v6i1.11333</dc:identifier>
	<dc:source xml:lang="en">Jurnal Kimia Valensi; Jurnal Kimia VALENSI Volume 6, No. 1, May 2020; 47-54</dc:source>
	<dc:source>2548-3013</dc:source>
	<dc:source>2460-6065</dc:source>
	<dc:language>eng</dc:language>
	<dc:relation>https://journal.uinjkt.ac.id/index.php/valensi/article/view/11333/pdf</dc:relation>
	<dc:rights xml:lang="en">Copyright (c) 2020 Iman Permana Maksum</dc:rights>
</oai_dc:dc>