Molecular Cloning of the cDNA Encoding Human Deoxyribonuclease II

A rapid amplification of cDNA ends method, using degenerate oligonucleotides based upon the N-terminal amino acid sequence of human hepatic deoxyribonuclease II (DNase II), allowed a novel cDNA encoding DNase II to be constructed from thyroid gland RNA. The composite nucleotide sequence (1593 bases)...

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Bibliographic Details
Published in:The Journal of biological chemistry 1998-01, Vol.273 (5), p.2610-2616
Main Authors: Yasuda, T, Takeshita, H, Iida, R, Nakajima, T, Hosomi, O, Nakashima, Y, Kishi, K
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Base Sequence
Chromosomes, Human, Pair 19
Cloning, Molecular
Deoxyribonuclease I - genetics
DNA, Complementary - genetics
Endodeoxyribonucleases - biosynthesis
Endodeoxyribonucleases - genetics
Escherichia coli - genetics
Gene Expression
Humans
Liver - enzymology
Molecular Sequence Data
Polymerase Chain Reaction
Recombinant Proteins - biosynthesis
RNA, Messenger - analysis
Sequence Analysis
Sequence Analysis, DNA
Sequence Homology, Amino Acid
Tissue Distribution
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Summary:A rapid amplification of cDNA ends method, using degenerate oligonucleotides based upon the N-terminal amino acid sequence of human hepatic deoxyribonuclease II (DNase II), allowed a novel cDNA encoding DNase II to be constructed from thyroid gland RNA. The composite nucleotide sequence (1593 bases) included an open reading frame of 1080 bases, which encoded a single polypeptide of 360 amino acids (signal peptide, 16; propeptide, 91; mature protein, 253). Although the sequence of DNase II showed no significant homology to other mammalian proteins, its cDNA structural organization resembled those of the lysosomal cathepsin families. The two parts of the cDNA corresponding to the propeptide and the mature protein were expressed in Escherichia coli , and the recombinant polypeptides thus obtained were strongly stained with an anti-DNase II antibody on Western blotting. DNase II is ubiquitously expressed in human tissues, and the DNase II gene ( DNASE2 ) was assigned to chromosome 19.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.273.5.2610