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All Journal Microbiology Indonesia Pharmacon Jurnal Bioteknologi & Biosains Indonesia (JBBI) ANNALES BOGORIENSES Berkala Penelitian Hayati Makara Journal of Science Jurnal Sains Materi Indonesia Jurnal Bioteknologi & Biosains Indonesia (JBBI) Annales Bogorienses Jurnal Bioteknologi & Biosains Indonesia
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Engineering Environmental Science Health Professions Immunology & microbiology Industrial & Manufacturing Engineering Materials Science & Nanotechnology Medicine & Pharmacology

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Journal : Makara Journal of Science

 1 
CLONING OF DENGUE VIRUS TYPE 3 (INDONESIAN STRAIN D3-1703) NON STRUCTURAL-1 GENE INTO pYES2/CT VECTOR Narita, Vanny; Widyanto, Rahma Micho; Pambudi, Sabar; Sudiro, Tjahjani Tjahjani
Makara Journal of Science Vol. 15, No. 2
Publisher : UI Scholars Hub

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Abstract

Dengue is an infectious disease caused by dengue virus. Dengue endemic region includes America, Western Pacific, Africa, East Mediterranian, and South East Asia including Indonesia. An early diagnostic system specific for Indonesia is needed to control dengue in Indonesia. In this research, cloning of Non Structural 1 (NS1) gene from dengue virus type 3 (Indonesian strain D3-1703) into pYES2/CT vector was performed. In the long run, NS1 recombinant protein will be expressed in Saccharomyces cerevisiae for diagnostic materials. Polymerase Chain Reaction (PCR) amplification of NS1 gene fragments were done with optimal annealing temperature at 55 ºC. NS1 gene fragment and pYES2/CT were cut by BamH I and Not I enzymes. The digested pYES2/CT was dephosphosrylated using Calf Intestine Alkaline Phospatase enzyme. Ligation with the vector:insert ratio of 1:12 and 1:20 resulted in 6 and 5 recombinant colony candidates respectively. Restriction enzyme and PCR verifications showed that 5 recombinant plasmids contained NS1 gene. Sequencing of the first 600 bp of one recombinant plasmid was performed. The blastn analysis showed that it had a 99% identity with dengue virus type 3 strain FW06. Finally, it was shown that NS1 clone within pYES2/CT was in the correct Open Reading Frame and ready to be expressed in S. cerevisiae.
Cloning and Expression of SCAMP3 in Escherichia coli BL21(DE3) with In Silico Sequence-Based Cancer Epitopes Prediction Rajagukguk, Selly Setiati; Pambudi, Sabar; Dwiranti, Astari; Utomo, Doddy Irawan Setyo; Bowolaksono, Anom
Makara Journal of Science Vol. 29, No. 1
Publisher : UI Scholars Hub

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Secretory carrier membrane protein 3 (SCAMP3) is a crucial membrane protein involved in intracellular vesicle traffick-ing and exocytosis. The SCAMP3 expression has been observed in diverse cancer types, such as melanoma, glioma, hepatocellular and breast cancer. Increased SCAMP3 expression has been reported in certain cancer cells relative to that in normal cells, suggesting the potential role of SCAMP3 in cancer development or progression. In this study, we successfully cloned and expressed SCAMP3 in Escherichia coli strain BL21(DE3). SCAMP3 was amplified and insert-ed directionally into the prokaryotic expression vector pET21d(+). The transformation of recombinant plasmid into E. coli BL21(DE3) cells were performed for the protein expression. SDS–PAGE and Western blotting were performed to detect the expression product induced by IPTG, which confirmed the presence of a recombinant pET21d(+)-SCAMP3 at 38-kDa protein weight. Bioinformatics analyses helped discover several possible epitopes distributed throughout the SCAMP3 protein sequence. These findings together serve as a basis for future biochemical and functional studies on this important membrane protein alongside immunotherapy research related to SCAMP3 as a cancer biomarker.
Co-Authors A.A. Ketut Agung Cahyawan W Abinawanto Abinawanto Alfan Danny Arbianto alfero Putra Iryanto ANGELINA GILL ANOM BOWOLAKSONO Arbianto, Alfan Danny Arbianto, Alfan Danny Arief Udhiarto ASRI SULFIANTI callista marsya CHURIYAH CHURIYAH Churiyah Churiyah DAMAI RIA SETYAWATI Dewi Esti Restiani DODDY IRAWAN SETYO UTOMO Dwiranti, Astari Etik Mardliyati Febriana Dwi Wahyuni feby feby HENI RACHMAWATI Ira Djajanegara Ira Djajanegara Ira Djajanegara Isma Nur Azzizah Lia Aprilia Nadya Stephanie nie nie Nina Artanti Nina Artanti Nora, Adri Novianti, Titta Nuralih Nuralih Nuralih Nuralih, Nuralih Okpri Meila Okpri Meila, Okpri putri mega utami Qonitah, Atikah Rahma Micho Widyanto Rahmana E. Kartasasmita Rahmana E. Kartasasmita Rajagukguk, Selly Setiati Ratno Nuryadi Restiani, Dewi Esti Restiani, Dewi Esti Retno Lestari Retno Lestari Retno Lestari roaslein putri Saraswati, Henny Seprianto, Seprianto SILMI RAHMANI Stephanie, Nadya Sudiro, Tjahjani Tjahjani Sukmadjaja Asyarie Sukmadjaja Asyarie Swasono R. Tamat Swasono R. Tamat Tarwadi Tarwadi Tarwadi, Tarwadi Tarwadi, Tarwadi Tika Widayanti Tika Widayanti TIKA WIDAYANTI Utomo, Doddy Irawan Setyo Vanny Narita Wayan Artama Whinie Lestari