Nur KUSMİYATİ, Nur HİDAYAT, Fatchiyah FATCHİYAH

Doğal Fermente Olan Sütteki Lactobacillus Spp.16S rRNA Geninin Kısmi Analizi

Amaç: Bu çalışmanın amacı doğal olarak fermente olan sütten Lactobacillus spp. türüne ait 16S rRNA ların gen sekansı yapmaktır. Materyal ve Metod: Kullanılan metodlar; alkali lizis yöntemiyle total DNA izolasyonu, spesifik primerlerle PCR amplifikasyonu ve amplifiye edilmiş DNA ların sekans analizlerinin yapılması şeklindedir. Elde edilen PCR ürünleri yaklaşık 585 bp uzunluğunda olup V1-V3 değişken bölgelerini içermektedir. Sekanslanan 16s rRNA daha sonra gen bankasıyla karşılaştırıldı. Bulgular: 16s rRNA geninin korunmuş bölgeleri; 189-358 ve 384-445 sırasıyla %95,9 ile %94,9 oranında korunurken, 16s rRNA geni totalde %78.8 oranında korunmuştur. Doğal sütün fermentasyonundan izole edilen bu 7 izolattan 2 tanesi L.Plantarum(benzerlik oranları %98.28 ve %97.78 diğer 5 izolattan 2"si L. Rhamnosus (benzerlik oranları (98.38 ve %97.51), diğer 2"si L. zeae (benzerlik oranları %97.36 ve % ve %98.38), biri ise L. case (benzerlik oranı%98.20) şeklinde tanımlanmıştır. Sonuç: Çalışmanın sonuçları test edilen izolatın Laktobailus genusuna ait olduğunu, fakat farklı türde sınıflandırıldığını göstermiştir.

Anahtar Kelimeler:

laktobacillus, doğal fermente süt, 16s rRNA, parsiyal analiz

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

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

Keywords:

16S rRNA gene, Lactobacillus, natural fermentation milk, partial analysis,

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Yazışma Adresi / Address for Correspondence: Dr. Fatchiyah Fatchiyah Dept of Biology, Faculty of Mathematics and Natural Sciences, Brawijaya University Malang, East Java, INDONESIA. e-mail: fatchiya@ub.ac.id, and fatchiya@gmail.com g eliş tarihi/received :05.09.2012 kabul tarihi/accepted:24.09.2013