Cloning, construction of prokaryotic expression vector and expression of Escherichia coli cytosine deaminase gene

Ren, S.; Jiang, H.; Gu, J.

Chinese Science Bulletin 43(3): 223-230


ISSN/ISBN: 1001-6538
Accession: 082454228

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Cytosine deaminase gene of Escherichia coli strain H-30 was cloned, and its initiation codon of 'GTG' was mutated to 'ATG' by PCR. Prokaryotic recombinant expression vector p BV220-Cd was constructed. Clone with high enzyme activity were selected by detecting their specific activity of cytosine deaminase. 5-Fc (5-FC, 5-fluorocytosine) could induce the lethal toxicity to cells containing active Cd gene. Dna sequence analysis indicated that there were 16 altered bases and 5 of them resulted in the alteration of amino acids in predicted peptide by comparing Dna sequence of the clone H-30-CD-11 with high enzyme activity with Cd gene reported in Gene Bank.