KARAKTERISASI GENETIK ISOLAT Bacillus thuringiensis BERDASARKAN GEN PENANDA 16s rRNA

Mega, Tyas Prihatin (2022) KARAKTERISASI GENETIK ISOLAT Bacillus thuringiensis BERDASARKAN GEN PENANDA 16s rRNA. Other thesis, Sekolah Tinggi Ilmu Kesehatan Nasional.

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Abstract

Bacillus thuringiensis is a rod-shaped bacterium, non-encapsulated, gram positive, motile with peritrichous flagella, and forms spores. This bacterium is a group of bacteria in the Cereus group. More than 82 Bt serovars were identified by flagellar antigen analysis (serotyping), however this test could not determine the relationship between variants. The purpose of this study was to determine the genetic diversity and relationbetween B. thuringiensis isolates based on the 16s rRNA gene, and to determine the pattern of diversity and relation of B.thuringiensis and other bacteria in the Cereus bacterial group. After End point PCR test with general 16s rRNA primer and sequencing by sanger sequencing, the DNA sequences were analyzed using MEGA X software. Alignment with references in the genebank was performed using Cluster W statistics and then the genetic distance was calculated. From the calculation of genetic distance, Salatiga isolates were divided into 3 large groups with genetic distance between isolates 0. 1st Group are Salatiga isolate 2,8,11,12,13,17 and 19. 2nd group are Salatiga isolate 6 and 7, lastly Salatiga isolate 9. Groups 1 and 2 have a genetic distance of 0.55%, groups 1 and 3 are 0.68 percent, groups 2 and 3 are 0.14%. From the calculation of the genetic distance, then constructed into a phylogenetic tree. From the construction of the phylogeny tree, salatiga isolates were divided into two clusters. Cluster 1, salatiga isolates 2,8,11,12,13,17 and 19 were close to B.thuringiensis serovar israelensis. Meanwhile, cluster 2 isolates 6,7 and 9 were close to B.thuringiensis serovar kurstaki. From the results of the analysis of secondary data on bacteria from the Cereus group in the genebank and the construction of the phylogenetic tree. 16s rRNA gene alone is not sufficient to differentiate and classify the bacterial group Cereus.

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