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Cukurova Med J. 2014; 39(1): 99-104


Partial Analysis 16S rRNA Gene in Lactobacillus spp. from Natural Fermented Milk

Nur Kusmiyati, Nur Hidayat, Fatchiyah Fatchiyah.

Abstract
Purpose: The aim of this research is to analysis 16S rRNA gene sequence of Lactobacillus spp. from natural fermented milk.
Material and Methods: The methods that used in this study are total DNA isolation using alkaline lysis, PCR amplification carried out using of specific primers, and identification of amplified DNA using sequencing. The result of PCR process is a nuclotide with approximately 585 bp length which contained V1-V3 variable regions. 16S rRNA gene sequence then compared and aligned with existing 16S rRNA data sequence from GenBank.
Results: The conserved region of 16S rRNA gene from nuclotide number 189-358 and 384-445 were 95,9% and 94,9% respectively, and the total conserved region of 16S rRNA gene is 78.8%. From total 7 isolates that isolated from natural fermented milk , 2 isolates were identified as L. plantarum with similarity value 98.28% and 97.78%, the other 5 isolates were identified as L. rhamnosus (2 isolates) with similarity value 98.38% and 97.51%, L. zeae (2 isolates) with similarity value 97.36% and 98.38%, and L. casei (1 isolate) with similarity value 98.20%.
Conclusion: These results indicate the test isolates are members of the genus Lactobacillus, but classified in different species

Key words: 16S rRNA gene, Lactobacillus, natural fermentation milk, partial analysis



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