Cloning, nucleotide sequence, and engineered expression of Thermus thermophilus DNA ligase, a homolog of Escherichia coli DNA ligase.
We have cloned and sequenced the gene for DNA ligase from Thermus thermophilus. A comparison of this sequence and those of other ligases reveals significant homology only with that of Escherichia coli. The overall amino acid composition of the thermophilic ligase and the pattern of amino acid substitutions between the two proteins are consistent with compositional biases in other thermophilic enzymes. We have engineered the expression of the T. thermophilus gene in Escherichia coli, and we show that E. coli proteins may be substantially removed from the thermostable ligase by a simple heat precipitation step.
Lauer
G
G
BioTechnica Diagnostics, Inc., Cambridge, Massachusetts 02138.
Rudd
E A
EA
McKay
D L
DL
Ally
A
A
Ally
D
D
Backman
K C
KC
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D011088:DNA Ligases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D004269:DNA, Bacterial / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis; D012689:Sequence Homology, Nucleic Acid; D013696:Temperature; D013824:Thermus / Q000201:enzymology / Q000235:genetics*
D004269:DNA, Bacterial; D011994:Recombinant Proteins; D011088:DNA Ligases