Applied and Environmental Microbiology · 1990 · 149 citations · 42 references
Life SciencesMicrobiologyMolecular BiologyEnzymologyGene ExpressionGenomicsPlasmid BiologyPlasmid EngineeringPlasmid GeneticsPlasmid GenomicsRecombinant Plasmid ConstructionThermostable EnzymesBacterial Open Reading FramesBacterial Plasmid ConstructionDna Sequence AnalysisGenomic Dna Library ConstructionGenomic Library ConstructionHomologyXylan DegradationXylanase Activity
A lambda recombinant bacteriophage coding for xylanase and beta-xylosidase activity has been isolated from a genomic library of the extremely thermophilic anaerobe "Caldocellum saccharolyticum." Partial Sau3AI fragments of the lambda recombinant DNA were ligated into pBR322. A recombinant plasmid with an insertion of ca. 7 kilobases of thermophilic DNA expressing both enzymatic activities was isolated. The location of the genes has been established by analyzing deletion derivatives, and the DNA sequence of 6.067 kilobases of the insert has been determined. Five open reading frames (ORFs) were found, one of which (ORF1; Mr 40,455) appears to code for a xylanase (XynA) which also acts on o-nitrophenyl-beta-D-xylopyranoside. Another, ORF5 (Mr 56,365), codes for a beta-xylosidase (XynB). The xynA gene product shows significant homology with the xylanases from the alkalophilic Bacillus sp. strain C125 and Clostridium thermocellum.
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Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
Ulrich K. Laemmli · Nature · 1970 · 251.6K citations