Molecular cloning of cDNA encoding Xenopus laevis deoxyribonuclease I.
A 1200-bp cDNA encoding Xenopus laevis deoxyribonuclease I (X. laevis DNase I) was constructed from the total RNA of a X. laevis pancreas using a rapid amplification of cDNA ends method. When the cDNA was transiently transfected into COS-7 cells, the recombinant polypeptide exhibited similar enzymological properties to those of the native pancreatic DNase I. The recombinant enzyme was considerably more labile than most other vertebrate DNase I enzymes. The X. laevis DNase I polypeptide was larger than any other known vertebrate DNase I, containing a unique Cys-rich stretch of 68 or 70 amino acid residues at the carboxyl terminus, and it had less well conserved binding sites for the Ca2+, G-actin and DNA, and two DNase I signature motifs. These alterations might account for its heat instability.
Hosomi
O
O
Department of Legal Medicine, Gunma University School of Medicine, Maebashi, Japan.
Yasuda
T
T
Takeshita
H
H
Nakajima
T
T
Nakashima
Y
Y
Mori
S
S
Mogi
K
K
Hanaoka
Y
Y
Kishi
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D018076:DNA, Complementary; D003850:Deoxyribonuclease I / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology; D010802:Phylogeny; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D014714:Vertebrates; D014982:Xenopus laevis / Q000235:genetics*
D018076:DNA, Complementary; D011994:Recombinant Proteins; D003850:Deoxyribonuclease I