Cloning, sequence analysis, and expression of the bacteriophage T4 cd gene.
The cd gene of bacteriophage T4, which encodes the enzyme deoxycytidylate deaminase, was isolated as a 1.9-kilobase DNA fragment and completely sequenced. The deduced amino acid sequence was found to be 193 residues long compared with 188 for the corresponding enzyme from bacteriophage T2. There were nine amino acid differences between the two enzymes in addition to a 5-residue insert near the carboxyl terminus of the T4 deaminase which was not present in the T2 deaminase. The cd-containing fragment also contained all of gene 31 (Nivinskas, R., and Black, L. W. (1988) Gene (Amst.) 73, 251-257) and thus precisely locates the two genes relative to one another within the T4 phage genomic map. Attempts to place the cd gene within a high expression vector have not been successful so far due to possible toxic effects of the gene product. However, placement of the gene within pUC18 resulted in a degree of expression which is about 10-20 times that found in T4-infected Escherichia coli. The enzyme was purified to homogeneity and found to possess properties similar to T2 phage deoxycytidylate deaminase.
Maley
G F
GF
Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509.
Duceman
B W
BW
Wang
A M
AM
Martinez
J
J
Maley
F
F
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D003844:DCMP Deaminase / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D015246:Deoxyribonuclease EcoRI; D015252:Deoxyribonucleases, Type II Site-Specific; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008969:Molecular Sequence Data; D009709:Nucleotide Deaminases / Q000235:genetics*; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D013604:T-Phages / Q000235:genetics*
D001426:Bacterial Proteins; D004279:DNA, Viral; D015246:Deoxyribonuclease EcoRI; C031851:endodeoxyribonuclease HaeII; C024404:BglII endonuclease; D015252:Deoxyribonucleases, Type II Site-Specific; D009709:Nucleotide Deaminases; D003844:DCMP Deaminase