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Enhanced Activity of Cytidine Deaminase by Gene Family Shuffling.  

Hong, Sik (Department of Microbiology, Kyungpook National University)
Kim, Kyung-Dong (Department of Microbiology, Kyungpook National University)
Song, Bang-Ho (Department of Biology Education, Kyungpook National University)
Jung, Kyung-Hwa (Genomics Laboratory, Amicogen Inc.)
Kim, Sa-Yeol (Department of Microbiology, Kyungpook National University)
Publication Information
Microbiology and Biotechnology Letters / v.30, no.4, 2002 , pp. 298-304 More about this Journal
Abstract
A family shuffling associating PCR-based and in vitro recombination and expression in Escherichia coli cdd mutant was carried out. Two cdd genes encoding cytidine deaminases (CDase) from thermophilic Bacillus caldolyticus and B. stearothermophilus were shuffled. Around 150 viable mutant colonies screened on AB minimal medium without uracil by E. coli cdd complementation were selected for cytidine deaminase assay and 4 candidates (SH1067, SH1077, SH1086, and SH1118) were chosen for the detailed study. The nucleotide sequence analyses of 4 selected mutants revealed that they have several point mutations and recombinations. Surprisingly, the SH 1067 showed 770 fold more specific CDase activity at $80^{\circ}C$ than that of T101 from parental B. stearothermophilus.
Keywords
Bacillus caldolyticus; B. stearothermophilus; cytidine deaminase; family shuffling.;
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Times Cited By SCOPUS : 1
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