ISOLASI DAN CLONING GEN PENYANDI α-AMILASE PADA ISOLAT BAKTERI AMILOLITIK, Bacillus sp. HS32

• Main Authors: , Almatin Puspa Dewi, , Dr. Yekti Asih Purwestri, S.Si., M.Si.
• Format: Theses and Dissertations NonPeerReviewed
• Published: [Yogyakarta] : Universitas Gadjah Mada 2014
• Subjects: ETD
• Online Access: https://repository.ugm.ac.id/134264/; http://etd.ugm.ac.id/index.php?mod=penelitian_detail&sub=PenelitianDetail&act=view&typ=html&buku_id=75332
• Institution: Universitas Gadjah Mada

Genes encoding α-amylase of amylolytic bacterial isolate, Bacillus sp. HS32 was successfully amplified by using Polymerase Chain Reaction (PCR) method produced band of 1.6 kb size and transformed into E. coli DH5α by using the zero blunt TOPO cloning vector. Selection of transformed cells was did in a selection medium containing kanamycin. Analysis of transformed cells by colony PCR method using primers Ami-F and Ami-R produced band of 1.6 kb and using M13 universal primers produced band of 1.8 kb size. Restriction Analysis of plasmid transformants by using restriction enzymes Not1 and BamH1 produced 2 bands, they were 3.5 kb (cloning vector) and 1.6 kb (insert gene). The results of sequencing of plasmid transformants and analysis of nucleotide similarity of α-amylase gene from isolates Bacillus sp. HS32 using the BLAST program on available database in the GenBank showed that the α-amylase gene from amylolytic bacterial isolate, Bacillus sp. HS32 had a similarity 96% with corresponding gene of Bacillus cereus NC7401.