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15 minutes 30 minutes acid sequence deduced acid sequence determined amino acid sequence amino terminal end ampicillin base pairs calf thymus DNA cellulase activity cellulase enzyme cellulase polypeptide cellulolytic cellulolytic microorganism Cellulomonas chilled on dry cloned cellulase gene Clostridium thermocellum coding codon usage cold 95 confirmed DNA sequence digested direction of transcription distilled water DNA was added dried under vacuum dry ice Eco R1 EDTA end-labeled DNA Eppendorf centrifuge Escherichia coli Ethanol Precipitations ethidium bromide Figure final pellet fusca strain yx hydrazine hydrogen bonds hydrophobic ice cold incubated at 37°C isolated Labeled DNA fragments minutes at 4°C Modification Reaction Seven molecular weight Nco l-Xma nucleotide sequence open reading frame pellet was dried plasmid DNA plasmid pD318 polyacrylamide gels protein re-precipitated region restriction endonuclease restriction enzyme sample secreted Sorvall Streptomvces subcloning experiments Thermomonospora DNA Thermomonospora fusca strain thymus Tris pH 8.0 tubes u.l of cold u.l of end-labeled