Cloning, Sequencing and Analyzing of 16S rRNA Gene from Inulin Hydrolyzing Bacteria

Putra, Ahadul and Azhar, Minda and Iryani, Iryani and Ahda, Yuni and Puspitasari, Fernita and Ihsanawati, Ihsanawati and Natalia, Dessy (2020) Cloning, Sequencing and Analyzing of 16S rRNA Gene from Inulin Hydrolyzing Bacteria. International Journal of Advances in Scientific Research and Engineering (ijasre), 6 (5). pp. 88-95. ISSN 2454-8006

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Abstract

Gene of 16S rRNA is used for molecular identification of bacteria. This research aims to analyze the sequences of 16S rRNA gene from inulin hydrolyzing bacteria of UKG isolate. The 16S rRNA gene was isolated by PCR using BacF1 and UniB1 primers. The gene was cloned to pGEM-T Easy vector with E.coli TOP10F as host cell. Recombinant DNA was sequenced using T7 and SP6 primers. The size of the nucleotide base of recombinant DNA of sequencing result using SP6 primer was 1167 bp, whereas using T7 primer was 1218 bp. The both of the sequences overlapped 760 bp. The size of 16S rRNA gene in the sequences was found 1501 bp. Recombinant DNA that was restricted using EcoR1 showed 3 bands on agarose gel. They were 3000 bp, 800 bp, and 700 bp. The position of the EcoR1 palindrome sequence on the 16S rRNA gene was at the 832-837. The 16S rRNA gene also has been known by HindIII, BamHI, BalI, HaeIII, and SmaI restriction enzymes.

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