• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.6
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• pp.699-707
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• 1995

Eurasian yellow perch (Perca fluviatilis) and American yellow perch (Perca flavescens) are known to be endemic species in Eurasia and North America, respectively. The presence of endemic species on each continent suggests their independent evolutionary history. However, because of the morphological similarity, distribution pattern, and only recent fossil record, their divergence time and speciation of the two Perca species has long been controversial. Here, from the comparison of the entire nucleotide sequences of cytochrome b gene, large genetic divergence between the two Perca species is observed although they are morphologically similar each other. Among 1,140 base pairs, interspecific nucleotide differences are found at 130 sites $(11.4\%)$. The differences varies with codon position, showing 22 sites in the first, 5 sites in the second, and 103 sites in the third codon position. Considering the types of nucleotide changes, transitional differences are much more than transversional differences and its ratio turned out to be 5.19. The estimated divergence time of the two Perca species indicates that they were separated each other approximately in the late Miocene period, which implies the long history of speciation. With comparison of the inferred amino acid sequences, strong structural and functional constraints which seem to be maintained by the highly conservative amino acid residues or protein regions, as found in other taxonomic groups of organisms, are also recognized in the cytochrome b of the fishes examined.

• Applied Biological Chemistry
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• v.31 no.3
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• pp.274-283
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• 1988

Aqualysin I is an alkaline serine protease which is secretet into the culture medium by Thermus aquaticus YT-1, an extreme thermophile. Aqualysin I was purified, and its partial amino acid sequence was determined. The gene encoding aqualysin I was cloned into E. coli using synthetic oligodeoxyribonucleotides as hybridization probes. The nucleotide sequence of the cloned DNA was determined. The primary structure of aqualysin I, deduced from the nucleotide sequenc, agreed with the determid amino acid sequences, including the $NH_2-$ and COOH terminal sequence of the tryptides derived from aqualysin I. Aqualysin I comprised 281 amino acid residues and its molecular mass was determined to be 28350. On alignment of the whole amino acid sequence, aqualysin I showed high sequence homology with the subtilisin type serine protease, and 43% identity with proteinase K, 37-30% with subtilisins and 34% with thermitase. Extremely high sequence identity was observed in the regions containing the active-site residues, corresponding to Asp32, His64 and Ser221 of subtilisin BPN'. Aqualysin I contains two disulfide bonds, Cys67-Cys99 and Cys163-Cys194, and these disulfide bonds seem to contribute to the heat stability of the enzyme. The determined positions of the twe disulfide bonds of aqualysin I agreed with those predicted previously on the basis of computer graphics of the crystallographic data for subtilisin BPN'. Therefore, these findings sugests that the three-dimensional structure of aqualysin I is similar to that of subtilisin BPN' Aqualysin I is produced as a lage precursor, which contains $NH_2-$ and COOH- terminal portions besides the mature protease sequence.