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Arsyam Mawardi
Cenderawasih University
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science Materials Science & Nanotechnology

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Kloning dan Analisis Bioinformatika Gen MSP1 Plasmodium falciparum Isolat Kota Jayapura Arsyam Mawardi; Leonardo E. Aisoi; Paula N. Lefaan
Jurnal Biologi Papua Vol 10, No 1 (2018)
Publisher : Jurusan Biologi FMIPA Universitas Cenderawasih

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (905.271 KB) | DOI: 10.31957/jbp.126

Abstract

Cloning gene involves the construction of a recombinant plasmid that inserted in a competent cell. On the other hand, genetic engineering requires bioinformatic analysis to be converted into tabulation and data interpretation. The study, titled "cloning block 2 MSP1 gene of Plasmodium falciparum isolate Jayapura city and bioinformatics analysis" is aimed to improve the technique of cloning the MSP1 gene of P. falciparum, initiated the creation of DH5α competent cells, ligations and transformations, plasmid isolation, confirmation the recombinant plasmid and able to perform bioinformatics analysis and construct phylogenetic tree. This study began with the manufacture of E. coli DH5α competent cells, MSP1 gene ligation in pJET1.2/blunt vector and transformation by using the heat shock transformation method, plasmid isolation of alkali lysis method, then plasmid confirmed by PCR and sequencing method, further sequence analysis and phylogenetic tree construction. The results showed that confirmation of MSP1 gene presence in pJET1.2/blunt with PCR was successful. From a total of 4 positive colonies grown in liquid culture, then isolated plasmid and confirmed with PCR obtained electroferogram bands with a size about 1049 bp indicates the presence of MSP1 gene in plasmid. Based on the results, cloning of MSP1 gene using pJET1.2/blunt cloning vector and competent cell E. coli DH5α has been successfully performed. Bioinformatics analysis of sequencing result and phylogenetic tree were constructed successfully with 2 clusters isolate of malaria patients from Jayapura city. Key words: Bioinformatics, cloning gene, heat shock transformation, MSP1, P. falciparum.
Analisis Perbandingan Kualitas Produk Amplikon Gen PMSA2 Antara Spesimen Spot Darah Kering dan Vena Arsyam Mawardi; Hendra K. Maury; Yustinus Maladan
Jurnal Biologi Papua Vol 12, No 1 (2020)
Publisher : Jurusan Biologi FMIPA Universitas Cenderawasih

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (326.927 KB) | DOI: 10.31957/jbp.949

Abstract

This study is aimed to analyze the comparative quality of PMSA2 gene amplicon product stability from two different specimen sources, spot specimens of dried blood and venous blood, as well as selecting the best storage method for specimens of blood samples. This research uses descriptive laboratory research methods. The research began with the process of sample preparation for dried blood spot and venous blood, each using Whatman 903 paper and vacuum tubes containing EDTA, isolating genomic DNA using KIT Zymo Research, amplification of PMSA-2 genes with PCR, detection of PCR products through electrophoresis, measurement of DNA concentration and absorbance, and data analysis. The results of this study are expected to be a source of information about the advantages of two specimen storage methods for clinical blood samples, as well as providing a clear description of the quality of each specimen storage method based on the quality of its amplicon products. The results showed that a total of ten medical samples of dried blood spot and ten venous blood were isolated from the genomic DNA of ten and nine, respectively. PMSA2 gene amplicons detected were seven in venous blood and six in dried blood spot. Venous blood specimens have sensitivity in detecting PMSA genes in samples with the highest value of 554 ng / μL and purity of 2,007 (WB7), and concentration of 550.2 and highest purity of 2,076 (WB10). Venous blood storage techniques using categorized vacuum tubes are effective in the detection of PMSA2 genes and have time efficiency in the process. From these results it was also concluded that the comparison analysis of amplicon products between venous blood specimens was better, more stable and efficient than dry blood spot specimens, thus recommending storage of venous blood specimens using vacuum tubes as the best storage method of blood sample specimens.
Co-Authors Hendra Kurniawan Maury Leonardo E. Aisoi Paula N. Lefaan Yustinus Maladan