KARAKTERISASI FRAGMEN 0,8 DAN 0,2 kb GEN PUTATIF LIPASE DARI BAKTERI Azospirillum sp. JG3

• Main Authors: , DILIN RAHAYU NATANINGTYAS, , Deni Pranowo, S.Si., M.Si.
• Format: Theses and Dissertations NonPeerReviewed
• Published: [Yogyakarta] : Universitas Gadjah Mada 2014
• Subjects: ETD
• Online Access: https://repository.ugm.ac.id/121875/; http://etd.ugm.ac.id/index.php?mod=penelitian_detail&sub=PenelitianDetail&act=view&typ=html&buku_id=61973
• Institution: Universitas Gadjah Mada

Characterization of 0.8 and 0.2 kb PCR fragment from bacterium Azospirillum sp. JG3 has been accomplished. This study is aimed to obtain nucleotide sequence of lipase gene fragment from the bacterium. PCR (Polymerase Chain Reaction) amplification was performed using primers that were designed based on the nucleotide sequence encoding acylglycerol lipase present in Azospirillum sp. B510 which has been published in genebank. The results of designed primer were synthesized and used for PCR using isolated DNA from bacterium Azospirillum sp. JG3 as template. The PCR conditions were: denaturation at 95 °C for 5 min, 35 cycles of denaturation at 95 °C for 30 s, annealing at 57 °C for 30 s, extension at 72 °C for 45 s, and final extension for 5 min. Two PCR fragments were isolated and purified for sequencing subsequently. The nucleotide sequences were analyzed by comparing the nucleotide sequence of lipase gene from Azospirillum sp. JG3 and others lipase genes from genebank. There were several candidate primers from design process and the selected primers were AzoF3 (5� GGA TCA CCT ATA CCC TCG TC 3�) and AzoR3 (5� CTT CAG GTC ACG CAA CAG 3�) which could amplify DNA template of this bacterium resulting 0.8 and 0.2 kb respectively. The sequence analysis of each PCR fragments showed that DNA sequences have 55.59% (0.8 kb) and 61.03% (0.2 kb) similarity to acylglycerol lipase DNA sequence of Azospirillum sp. B510. Therefore the amplified 0.8 and 0.2 kb fragments could be part of lipase putative gene.