Detail Cloning and Sequencing of Thermostable Mannanase Gene from Geobacillus stearothermophilus L-07 Bibliografi Author: Theosophus, Yohanes ; Suwanto, Antonius (Advisor) Topik: Geobacillus Stearrothermophilus L-07 Bahasa: (EN )     Penerbit: Fakultas Teknobiologi Unika Atma Jaya (Faculty of Biotechnology Atma Jaya Catholic University of Indonesia)     Tempat Terbit: Jakarta    Tahun Terbit: 2006     Jenis: Theses - Undergraduate Thesis Fulltext: Yohanes Theosophus's Undergraduated Theses.pdf (663.64KB; 18 download) Ketersediaan Perpustakaan Pusat (Semanggi) Nomor Panggil: FTB-015 Non-tandon: tidak ada Tandon: 1  Lihat Detail Induk Abstract A 2.3-kb BamHI-HindIII fragment containing putative mannanase gene from Geobacillus stearothermophilus L-07 has been identified through Southern hybridization analysis. This DNA fragment was ligated to pAS900 vector for Inverse-PCR analysis and DNA sequencing. The Inverse-PCR amplicon sequences showed high similarity to the genes, manF and estA, from Bacillus stearothermophilus ATCC 12016 (accession no. AF038547). Employment of both methods demonstrated that the DNA band carried a truncated ß-mannanase gene on its upstream and an initial part of esterase gene on the downstream. A pair of new primers was designed to amplify a specific ß-mannanase full-length gene (manL) CDS from strain L-07 and resulting 2.0-kb DNA fragment. The fragment was ligated to pGEM®-T Easy Vector and transformed into E. coli DH5a. The plasmids from white colonies were isolated and digested with restriction endonuclease EcoRI to verify the presence of insert DNA. The single- and double-digested of one of the recombinant plasmids showed that the insert DNA was truly a manL full-length gene since the electropherogram was identical to the manF restriction endonuclease analysis. The insert DNA was subsequently sequenced and partial alignment showed that this sequence has 98% identities to the manF. Opini Anda Klik untuk menuliskan opini Anda tentang koleksi ini! Lihat Sejarah Pengadaan  Konversi Metadata   Kembali

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