Identifikasi spesies Bacillus sp. K2Br5 dengan analisis urutan Gen 16s rRNA

Tafiani, Nurul (2018) Identifikasi spesies Bacillus sp. K2Br5 dengan analisis urutan Gen 16s rRNA. Diploma thesis, UIN Sunan Gunung Djati Bandung.

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Abstract

INDONESIA: Identifikasi diperlukan untuk mengetahui spesies bakteri. Identifikasi bakteri dapat dilakukan secara fenotip maupun genotip. Namun, identifikasi secara fenotip memiliki kelemahan yakni sering terjadi kesalahan dalam menentukan spesies dan galur bakteri. Oleh karena itu, diperlukan identifikasi secara genotip melalui identifikasi urutan gen 16s rRNA. Penelitian ini bertujuan untuk mengidentifikasi spesies bakteri dengan kode K2Br5 yang telah diisolasi dari kawasan tanah Karst yang kemudian teridentifikasi secara fenotip memiliki genus Bacillus. Pada penelitian ini dilakukan analisis secara genotip dengan langkah awal mengisolasi DNA kromosom dari bakteri Bacillus sp. K2Br5. Selanjutnya dilakukan proses amplifikasi dengan metode PCR untuk memperoleh fragmen DNA 16s rRNA menggunakan primer maju universal BactF1 dan primer mundur UniB1. Fragmen gen 16s rRNA yang diperoleh selanjutnya disekuensing dengan metode dideoksi Sanger untuk mengetahui urutan nukleotida. Urutan nukleotida hasil sekuensing dimasukkan kedalam program BLAST untuk mengetahui homologinya dengan urutan nukleotida fragmen gen 16s rRNA bakteri lain yang telah terdata pada bank gen. Hasil BLAST menunjukkan bahwa bakteri Bacillus sp. K2Br5 memiliki kemungkinan berkerabat dengan Paraclostridium bifermentans yang memiliki nama lain Clostridium bifermentans, Martellillus bifermentans, Bacillus biferementans, Bacillus bifermentans sporogenes, dan Bacillus centrosporogenes. ENGLISH: Identification is needed to determine bacterial species. Bacterial identification can be done either phenotypically or genotypically. However, identification in a phenotype has the disadvantage of frequent errors in determining bacterial species and strains. Therefore, genotive identification is needed through identification of 16s rRNA gene sequences. This study aimed to identify bacterial species coded K2Br5 which had been isolated from the Karst soil area which was then identified phenotically as having the genus Bacillus. In this study genotypic analysis was carried out with the initial step of isolating chromosomal DNA from the bacterium Bacillus sp. K2Br5. The amplification process was then carried out by the PCR method to obtain the 16s rRNA DNA fragment using the advanced universal primer BactF1 and the UniB1 reverse primer. The 16s rRNA gene fragments obtained were then sequenced by the deeoxy Sanger method to determine nucleotide sequences. Sequencing nucleotide sequences were entered into the BLAST program to find out the homology with the nucleotide sequences of 16s rRNA gene fragments of other bacteria that had been recorded in the gene bank. BLAST results show that Bacillus sp. K2Br5 has the possibility of being related to Paraclostridium bifermentans which have other names Clostridium bifermentans, Martellillus bifermentans, Bacillus biferementans, Bacillus bifermentans sporogenes, and Bacillus centrosporogenes.

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