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Ligasi dan Transformasi Gen MSP1 Plasmodium falciparum Penyebab Malaria di Kota Jayapura Mawardi, Arsyam; Ramandey, Euniche R.P. F.
Majalah Kedokteran Bandung Vol 49, No 4 (2017)
Publisher : Faculty of Medicine, Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (484.5 KB) | DOI: 10.15395/mkb.v49n4.1138

Abstract

MSP1 merupakan protein yang antigenik dan paling banyak diekpresikan pada permukaan merozoit ketika menginfeksi eritrosit pasien malaria sehingga banyak dikembangkan untuk desain terapi vaksin. Proses ligasi dan transformasi gen MSP1 merupakan upaya penggandaan gen untuk menghasilkan produk yang sama ketika diekspresikan. Penelitian ini bertujuan mengkloning gen MSP1 P. falciparum  dari pasien malaria tropika di Jayapura menggunakan vektor pJET1.2/blunt dan sel kompeten E. coli DH5 sehingga didapatkan perbanyakan plasmid rekombinan yang mengandung gen MSP1. Darah yang positif mengandung P. falciparum diproses secara molekuler, diawali tahapan isolasi DNA genom, amplifikasi dengan teknik PCR, ligasi ke dalam vektor pJET1.2/blunt dan ditransformasi pada E. coli DH5α dengan metode Heat Shock Transformation, diakhiri dengan konfirmasi PCR untuk memastikan tersisipkannya gen blok 2 MSP1. Hasil penelitian menunjukkan bahwa konfirmasi keberadaan gen MSP1 dalam pJET1.2/blunt dengan PCR berhasil dilakukan. Dari total 10 koloni positif  yang ditumbuhkan dalam kultur cair, kemudian diiisolasi plasmid dan dikonfirmasi dengan PCR diperoleh pita hasil elektroferogram dengan ukuran sekitar 1049 bp yang menunjukan gen MSP1 dalam plasmid. Berdasar atas hasil tersebut, kloning gen MSP1 menggunakan vektor kloning pJET1.2/blunt dan sel kompeten E. coli DH5a telah berhasil dilakukan. Kata kunci: Heat Shock, ligasi, MSP1, P. falciparum, transformasi Malaria-causing MSP-1 Plasmodium falciparum Ligation and Transformation in Jayapura CityMSP1 is the most antigenic and expressed protein on merozoite surface when it infects the erythrocytes of malaria patients which leads to its use for vaccine therapy design development. The ligation and transformation process of the MSP1 gene is a gene duplication attempt for producing  the same product during expression. This study aimed to clone P. falciparum MSP-1 gene from tropical malaria patients in Jayapura using pJET1.2/blunt vectors and E. coli DH5a competent cells, to get the recombinant plasmid propagation of MSP1 gene. Blood that was positive for P. falciparum was molecularly processed, starting with genomic DNA isolation and then followed by PCR amplification, ligation into pJET1.2/blunt vector, and transformation into E. coli DH5α using the heat shock transformation method. The process was ended with PCR confirmation to confirm MSP1 gene insertion. The results showed that the presence of the  MSP1 gene in pJET1.2/blunt was successfully confirmed through PCR. From a total of 10 positive colonies grown in liquid culture,  plasmid was isolated. Electropherogram result presented bands  of about 1049bp, indicating the presence of the MSP1 gene in plasmid. Hence, MSP1 gene cloning using pJET1.2/blunt cloning vector and competent cell E. coli DH5α has been successfully performed. Key words: Heat shock, ligation, MSP-1, P. falciparum, transformation  
Sequence analysis of 18SrDNA gene from sagoplast degrading fungi Tri Gunaedi; Arsyam Mawardi
JURNAL PENELITIAN BIOLOGI BERKALA PENELITIAN HAYATI Vol 26 No 2 (2021): June 2021
Publisher : The East Java Biological Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23869/bphjbr.26.2.20214

Abstract

The bioplastic can be made from sago flour and known as sagoplast. It was widely known that for making bioplastic, the addition of acetic acid and glycerol are needed. Products that are air-dried are easy to grow fungi within a few weeks. This makes the basis for researchers to undestand more about the character and identity of the sagoplast degrading fungi. Characterization and identification were carried out by observed morphology and analyzing the 18SrDNA gene sequence of fungal isolates that had grown on the sagoplast. Fungal isolates morphology showed yellowish-orange color with white thread-like mycelia and a blackish brown mace with white thread-shaped mycelia. These characters of fungal morphology that similar with Aspergillus. The gene sequences of the fungal isolates were aligned with reference gene sequences of the fungi obtained from the Gen Bank of the National Center for Biotechnology Information (NCBI). Sequence data analysis was performed by using the Clustal X program to determine the kinship and taxonomy of the fungal isolates that able to degrade sagoplast. The result showed that two fungal isolates, DFSP.J1 and DFSP.J4, were found and demonstrated their ability for degrading sagoplast. Isolate DFSP.J1 is related to Aspergillus flavus strain PSU2 LC127086.1, while isolate DFSP.J4 is related to Aspergillus niger IFO4033 D63697.1.