Molecular, biochemical and immunological analyses of canine pancreatic DNase I |
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Authors: | Kaneko Yasushi Takeshita Haruo Mogi Kouichi Nakajima Tamiko Yasuda Toshihiro Itoi Masako Kuwano Hiroyuki Kishi Koichiro |
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Institution: | Department of Legal Medicine and Molecular Genetics, Gunma University, Graduate School of Medicine, Maebashi, Gunma 371-8511. |
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Abstract: | The DNase I from canine pancreas was purified 260-fold to electrophoretic homogeneity with a 35% yield using three-step column chromatography. The activity of the purified enzyme was completely inhibited by 20 mM EDTA, an antibody specific to the purified enzyme and G-actin. A 1,373-bp cDNA encoding canine DNase I was constructed from the total canine pancreatic RNA using a rapid amplification of cDNA ends method, followed by sequencing. The mature canine DNase I protein was found to consist of 262 amino acids. A survey of DNase I in 13 different canine tissues revealed the highest levels of both DNase I enzyme activity and gene expression in the pancreas; therefore, the canine DNase I is of the pancreatic type. Phylogenetic and sequence identity analyses, studies of immunological properties and the tissue-distribution patterns of DNase I indicated that the canine enzyme is more closely related to the human DNase I than to other mammalian DNases I. Therefore, canine DNase I is found to be one of the best substitutes in studies of human DNase I. |
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