Open AccessIdentification of the bacterium isolate from Mackerel Fish (Rastrelliger sp.) using 16S rRNA Gene
Author(s) -
Yulia Sari Ismail,
. Febriani,
Cut Yulvizar,
Rahayu Ramadhani
Publication year - 2019
Publication title -
iop conference series. earth and environmental science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.179
H-Index - 26
eISSN - 1755-1307
pISSN - 1755-1315
DOI - 10.1088/1755-1315/293/1/012043
Subject(s) - biology , genbank , 16s ribosomal rna , homology (biology) , phylogenetic tree , bacillus megaterium , ribosomal rna , genetics , gene , sequence analysis , microbiology and biotechnology , bacteria
The use of probiotics in aquaculture can improve the quality and quantity of the fishes. One of potential probiotic bacterium from gastrointestine of mackerel fish (Rastrelliger sp.) isolated in this research were LUI-04. This study aimed to determine the species of bacterium isolate using nucleotide sequence homology compared to other microorganisms in GenBank. The 16S rRNA genes of LUI-04 isolate were analyzed. The results of electrophoresis showed the DNA band had a size of 1 500 bp for amplification using Bact-27F and Uni-1492R primers, and about 250 bp to 500 bp with the Com-IF and Com-2R primers. The sequence of nucleotides was determined by Sanger method. The 16S rRNA sequence of the isolate and other bacteria in GenBank were compared with the Basic Alignment Search Tool (BLAST) to know the homology. Then, all of the sequences were aligned by ClustalW, and the phylogenetic tree was reconstructed with Neighbor Joining method using bootstrap analysis of 1 000 replicates. The results showed that LUI-04 had a homology of 99 % and the bootstrap value percentage of 86 % with Bacillus aryabhattai B8W22 and four strains of Bacillus megaterium (strain QM B1551, NBRC15308, strain IAM 13418 and ATCC 14581). Therefore, the LUI-04 isolate belongs to Bacillus megaterium species.