CLONING AND EXPRESSION OF Plasmodium falciparum LACTATE DEHYDROGENASE (PfLDH) IN Escherichia coli BL21(DE3)

Acta Biochimica Indonesiana

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Title CLONING AND EXPRESSION OF Plasmodium falciparum LACTATE DEHYDROGENASE (PfLDH) IN Escherichia coli BL21(DE3)
 
Creator Masduki, Fifi Fitriyah
Hotimah, Yeni
Rani, Rosalia
Mawardi, Arsyam
Ramandey, Euniche R.P.F.
Fibriani, Azzania
Suhandono, Sony
 
Description Background: Immediate and accurate diagnosis of malaria is essential for effective control of this disease. Immunochromatographic based rapid diagnostic tests (RDTs)  are economical, simple to perform, and provide results in a relative short time, can be useful to assist effective management of malaria. The commercially available malaria RDT in Indonesia is still imported. Therefore, an effort to produce malaria RDT independently is necessary. One of the biomarkers used in RDTs is Plasmodium lactate dehydrogenase pLDH. The production and accumulation of pLDH during asexual stage or blood-stage in all human infected malaria parasites can be used to indicate parasites viability, which is correlated with the number of parasites present in the plasma of infected patients.  Objective: The aim of this research is to produce recombinant PfLDH in Escherichia coli BL21(DE3).Methods: PfLDH gene was cloned into pET30a expression vector to obtain a 6.2 kbp recombinant plasmid pET30a-PfLDH. E. coli BL21(DE3) was transformed with pET30a-PfLDH using the heat shock method. Then, E. coli BL21(DE3)- pET30a-PfLDH was cultured in LB broth containing 50 mg/mL kanamycin and was induced by 1mM IPTG at 37oC.Results: SDS-PAGE and Western Blot analysis showed that recombinant PfLDH was expressed with molecular mass ~30 kDa.Conclusion: Recombinant PfLDH is expressed in E. coli BL21(DE3) and can be used in further research for producing rPfLDH as a biomarker for malaria RDT development.
 
Publisher PP PBBMI
 
Contributor Fifi Fitriyah Masduki, ITB
Yeni Hotimah, ITB
Rosalia Rani, ITB
Arsyam Mawardi, Universitas Cendrawasih
Euniche R.P.F. Ramandey, Universitas Cendrawasih
Azzania Fibriani, ITB
Sony Suhandono, ITB
 
Date 2020-02-08
 
Type info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Peer-reviewed Article
 
Format application/pdf
 
Identifier https://pbbmi.org/jurnal/index.php/ActaBioIna/article/view/39
10.32889/actabioina.v2i2.39
 
Source Acta Biochimica Indonesiana; Vol 2, No 2 (2019): ACTA BIOCHIMICA INDONESIANA
2654-3222
2654-6108
 
Language eng
 
Relation https://pbbmi.org/jurnal/index.php/ActaBioIna/article/view/39/pdf_1
https://pbbmi.org/jurnal/index.php/ActaBioIna/article/downloadSuppFile/39/2
 
Rights Copyright (c) 2020 Fifi Fitriyah Masduki
https://creativecommons.org/licenses/by-nc/4.0
 

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