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AssigneeTrigen Limited
InventorsHun-Chi Lin, Shau-Ping Lei
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Genetically engineered E. coli carry vectors containing inserts that code for Clostridium histolyticum collagenase. These inserts code for: (a) a form of collagenase having a molecular weight of about 68,000 daltons in the essential absence of larger forms of collagenase; (b) the 68 kd form of collagenase and a fusion polypeptide consisting of the collagenase protein fused to at least a portion of the .beta.-galactosidase protein of E. coli; or (3) the 68 kd form of collagenase and polypeptides of molecular weight of from above about 68,000 daltons to about 100,000 daltons and having the enzymatic activity of C. histolyticum collagenase as indicated by digestion of .sup.3 H-acetylated collagen and by specific inhibition by 1,10-phenanthroline plus EDTA. The collagenase genes in the transformed E. coli are expressed efficiently in the transformed cells to yield enzymatically active and immunologically cross-reactive collagenase. In particular, the 68 kd form of collagenase is resistant to autocatalytic degradation and is stable to long-term storage. Genetically engineered collagenase, especia The research reported in this application in support of the invention herein received assistance from a grant provided by the Department of Health & Human Services, Public Health Service, National Institute of Diabetes, Digestive and Kidney Disease, Grant No. N43-DK-8-2239.
An isolated and purified recombinant DNA fragment consisting essentially of DNA derived from Clostridium histolyticum coding solely for a polypeptide having the enzymatic activity of C. histolyticum collagenase, the polypeptide having a molecular weight of about 68,000 daltons.
The DNA fragment of claim 1 capable of being transcribed to yield mRNA, the mRNA being capable of being translated to yield a polypeptide of about 68,000 daltons molecular weight and having collagenase activity, the polypeptide being distinguishable by the heterologous expression of the sequence of claim 3 from the endogenous production by C. histolyticum of mutliple forms of collagenase by the essential absence of those forms of C. histolyticum collagenase having molecular weights of above about 70,000 daltons determined by the expression of the C. histolyticum genomic coding sequence.
The DNA fragment of claim 1 capable of being transcribed to yield mRNA, the mRNA being capable of being translated to yield a peptide displaying the antigenicity of C. histolyticum collagenease, the polypeptide being distinguishable by the heterologous expression of the sequence of claim 3 from the endogenous production by C. histolyticum of multiple forms of collagenase by the essential absence of those forms of C. histolyticum collagenase having molecular weights of above about 70,000 daltons determined by the expression of the C. histolyticum genomic coding sequence.
Dependencies are inferred from the claim text.