Characterization, cloning and expression of the 67-kDA annexin from chicken growth plate cartilage matrix vesicles.
Analysis of a 67 kDa lipid-dependent Ca(2+)-binding protein from avian matrix vesicles revealed amino acid sequences homologous to mammalian annexin VI. PCR methods were used to identify a clone from an avian cDNA library that contained a full length copy of the 67-kDa annexin cDNA. This was restriction mapped, subcloned and sequenced. The cDNA sequence of the open reading frame showed 70 percent identity to that of murine annexin VI; the predicted amino acid sequence, 78 percent identity. There was no homology in the 5'- and 3'-untranslated regions. A plasmid was constructed that overexpresses the intact chicken 67-kDa matrix vesicle annexin in E. coli DH5 alpha in high yield; the physicochemical properties and the amino terminal sequence of the expressed protein exactly matched those of the native protein.
Cao
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Department of Chemistry and Biochemistry, University of South Carolina, Columbia 29210.
Genge
B R
BR
Wu
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Buzzi
W R
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Showman
R M
RM
Wuthier
R E
RE
D000595:Amino Acid Sequence; D000818:Animals; D017317:Annexin A6 / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D001483:Base Sequence; D002645:Chickens; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000096:biosynthesis; D004926:Escherichia coli; D015870:Gene Expression; D006132:Growth Plate / Q000378:metabolism*; D008322:Mammals; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009838:Oligodeoxyribonucleotides; D016366:Open Reading Frames; D015388:Organelles / Q000378:metabolism*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping
D017317:Annexin A6; D018076:DNA, Complementary; D009838:Oligodeoxyribonucleotides