Purification of triosephosphate isomerase and isolation of its gene from the mosquito Culex tarsalis.
The enzyme triosephosphate isomerase (TPI) was purified to homogeneity from the mosquito Culex tarsalis. Anti-C. tarsalis TPI antibodies cross-reacted with TPIs from other organisms but bands on western blots were most intense with proteins from closely related Dipterans. Using a degenerate primer corresponding to the amino-terminal sequence of the protein in a polymerase chain reaction (PCR), a cDNA corresponding to the TPI gene (Tpi) was isolated and sequenced. Subsequently, a genomic sequence including 305 bp to the 5'-end of the coding sequence was obtained. Comparison of C. tarsalis Tpi to that of Drosophila melanogaster revealed that although the two genes had little similarity in the intron and 5' flanking sequences, they were highly similar (73% identity) in their coding sequence. The rate of synonymous substitution in insect genes may be slower than that of vertebrates, but the nonsynonymous substitution rate, and hence the rate of TPI evolution, appears to be faster in insects than in vertebrates.
Whyard
S
S
Department of Biology and Insect Biotech Canada, Queen's University, Kingston, Ontario.
Tittiger
C
C
Walker
V K
VK
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D005075:Biological Evolution; D003001:Cloning, Molecular; D003465:Culex / Q000201:enzymology* / Q000235:genetics; D004247:DNA / Q000032:analysis; D004331:Drosophila melanogaster / Q000201:enzymology / Q000235:genetics; D005260:Female; D017344:Genes, Insect; D008297:Male; D008969:Molecular Sequence Data; D014305:Triose-Phosphate Isomerase / Q000235:genetics / Q000302:isolation & purification*
D004247:DNA; D014305:Triose-Phosphate Isomerase