Cloning of cDNAs encoding porcine and human DNase II.
We report the molecular cloning of cDNAs encoding porcine and human DNase II and the genomic structure of the human DNase II gene. The full length cDNAs for porcine and human DNase II were isolated by polymerase chain reaction on the basis of amino acid sequences determined for the tryptic peptides of porcine liver DNase II. The porcine and human cDNAs contain 1095 and 1083 bp open reading frames, respectively, and encode 364 and 360 amino acid proteins with calculated molecular masses of 40,157 and 39,555, respectively. The amino acid sequencing of purified porcine DNase II reveals two N-termini with corresponding sequences present within the same open reading frame, suggesting proteolytic processing for the covalently bonded subunit structure of DNase II. Northern blot analysis demonstrated that a single transcript of 2.0 kb mRNA coding for DNase II is ubiquitously expressed in human tissues. A database search revealed that the human genomic sequence of chromosome 19p13.2 contains the DNase II gene. Characterization of the genomic sequence showed that the DNase II gene consists of six exons separated by five introns whose splice acceptor/donor sites agree with the GT/AG rule.
Shiokawa
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Department of Biochemistry, Faculty of Pharmaceutical Sciences, Science University of Tokyo, Japan.
Tanuma
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002888:Chromosomes, Human, Pair 19 / Q000235:genetics; D003001:Cloning, Molecular; D004706:Endodeoxyribonucleases / Q000737:chemistry*; D005091:Exons / Q000235:genetics; D006801:Humans; D007438:Introns / Q000235:genetics; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D010449:Peptide Mapping; D016133:Polymerase Chain Reaction; D011499:Protein Processing, Post-Translational / Q000502:physiology; D012326:RNA Splicing / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013552:Swine
D012333:RNA, Messenger; D004706:Endodeoxyribonucleases; C019329:deoxyribonuclease II