Gene serum opacity factor
AssigneeThe Rockefeller University
InventorsVincent A. Fischetti, Jasna Rakonjac, John Robbins
Life of this patent
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Abstract
Methods and compositions are provided for cloning and expression of serum opacity factor of Streptococcus pyogenes genes. The portion produced by the recombinant DNA techniques described herein may be employed in qualitative and quantitative testing for high density lipoprotein, as a fibronectin binding factor and for the regulation of high density lipoprotein in a mammal. The gene may further be employed as a molecular probe for accurate identification of opacity factors from various strains of Streptococcus pyogenes.
Claims19
A process for producing a polypeptide having apolipoproteinase activity which comprises culturing a prokaryotic unicellular organism containing a recombinant plasmid comprising a DNA sequence coding for said polypeptide and capable of being replicated, transcribed and translated in the unicellular organism, and isolating said polypeptide from the culture, said polypeptide being encoded by a sequence selected from the group consisting of:
- (1) the DNA sequences of FIG. 4 (SEQ ID NO. 2);
- (2) DNA sequences that hybridize to the DNA sequence of (1) under high stringency hybridization conditions;
- and
- (3) DNA sequences that encode an amino acid sequence encoded by the DNA sequences of (1) or (2), and enzyme active segments thereof.
A process for preparing a prokaryotic unicellular organism having a DNA sequence coding for a polypeptide having apolipoproteinase activity which comprises introducing a recombinant plasmid comprising a DNA sequence coding for said polypeptide and capable of being replicated, transcribed and translated in the unicellular organism, said sequence selected from the group consisting of:
- (1) the DNA sequence of FIG. 4 (SEQ ID NO. 2);
- (2) DNA sequence that hybridize to the DNA sequence of (1) under high stringency hybridization conditions;
- and
- (3) DNA sequences that encode an amino acid sequence encoded by the DNA sequences of (1) or (2), and enzyme active segments thereof.
A purified DNA sequence coding for a polypeptide having apolipoproteinase activity, said sequence being selected from the group consisting of:
- (1) the DNA sequence of FIG. 4 (SEQ ID NO. 2);
- (2) DNA sequence that hybridize to the DNA sequence of (1) under high stringency hybridization conditions;
- and
- (3) DNA sequences that encode an amino acid sequence encoded by the DNA sequences of (1) or (2), and enzyme active segments thereof.
A purified DNA probe capable of binding to a DNA sequence coding for a polypeptide having apolipoproteinase activity or any portion of said sequence having such activity, said sequence being selected from the group consisting of:
- (1) the DNA sequence of FIG. 4 (SEQ ID NO. 2);
- (2) DNA sequence that hybridize to the DNA sequence of (1) under high stringency hybridization conditions;
- and
- (3) DNA sequences that encode an amino acid sequence encoded by the DNA sequences of (1) or (2), and enzyme active segments thereof.
A diagnostic test for high density lipoprotein in a body fluid which comprises:
- (a) contacting said body fluid with a polypeptide having apolipoproteinase activity, said polypeptide being encoded by one of the following sequences:
- 1. the DNA sequence of FIG. 4 (SEQ ID NO:2);
- 2. DNA sequences that hybridize to the DNA sequences of (1) under high stringency hybridization conditions;
- and
- 3. DNA sequences that encode an amino acid sequence encoded by the DNA sequences of (1) or (2);
- (b) detecting cleavage of apoprotein A1;
- and
- (c) correlating said cleavage with the presence of high density lipoprotein.
Dependencies are inferred from the claim text.