Hyaluronan synthase gene and uses thereof
First Claim
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1. A purified nucleic acid segment comprising a coding region encoding enzymatically active hyaluronate synthase.
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Abstract
The present invention relates to a nucleic acid segment having a coding region segment encoding enzymatically active Streptococcus equisimilis hyaluronate synthase (seHAS), and to the use of this nucleic acid segment in the preparation of recombinant cells which produce hyaluronate synthase and its hyaluronic acid product. Hyaluronate is also known as hyaluronic acid or hyaluronan.
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Citations
59 Claims
- 1. A purified nucleic acid segment comprising a coding region encoding enzymatically active hyaluronate synthase.
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4. A purified nucleic acid segment having a coding region encoding enzymatically active hyaluronate synthase, wherein the purified nucleic acid segment is capable of hybridizing to the nucleotide sequence of SEQ ID NO:
- 1.
- 5. A purified nucleic acid segment having a coding region encoding enzymatically active hyaluronate synthase, wherein the purified nucleic acid segment has semiconservative or conservative amino codon acid changes when compared to the nucleotide sequence of SEQ ID NO:
- 6. A recombinant vector selected from the group consisting of a plasmid, cosmid, phage, or virus vector and wherein the recombinant vector further comprises a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase.
- 11. A recombinant host cell, wherein the recombinant host cell is a prokaryotic cell transformed with a recombinant vector comprising a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase.
- 17. A recombinant host cell, wherein the recombinant host cell is a eukaryotic cell transfected with a recombinant vector comprising a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase.
- 23. A recombinant host cell, wherein the recombinant host cell is electroporated to introduce a recombinant vector into the recombinant host cell, wherein the recombinant vector comprises a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase.
- 29. A recombinant host cell, wherein the recombinant host cell is transduced with a recombinant vector comprising a purified nucleic acid segment having a coding region encoding enzymatically active Streptococcus equisimilis hyaluronan synthase.
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35. A purified composition, wherein the purified composition comprises an enzymatically active hyaluronan synthase polypeptide.
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36. A purified composition, wherein the purified composition comprises a polypeptide having an amino acid sequence in accordance with SEQ ID NO:
- 2.
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37. A method for detecting a DNA species, comprising the steps of:
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obtaining a DNA sample;
contacting the DNA sample with a purified nucleic acid segment in accordance with SEQ ID NO;
1;
hybridizing the DNA sample and the purified nucleic acid segment thereby forming a hybridized complex; and
detecting the complex.
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38. A method for detecting a bacterial cell that expresses mRNA encoding Streptococcus equisimilis hyaluronan synthase, comprising the steps of:
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obtaining a bacterial cell sample;
contacting at least one nucleic acid from the bacterial cell sample with purified nucleic acid segment in accordance with SEQ ID NO;
1;
hybridizing the at least one nucleic acid and the purified nucleic acid segment thereby forming a hybridized complex; and
detecting the hybridized complex, wherein the presence of the hybridized complex is indicative of a bacterial strain that expresses mRNA encoding Streptococcus equisimilis hyaluronan synthase.
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39. A method for producing hyaluronic acid, comprising the steps of:
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introducing a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase into a host organism, wherein the host organism contains nucleic acid segments encoding enzymes which produce UDP-GlcNAc and UDP-GlcA;
growing the host organism in a medium to secrete hyaluronic acid; and
it recovering the secreted hyaluronic acid. - View Dependent Claims (40, 41, 42, 43)
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- 44. A pharmaceutical composition comprising a preselected pharmaceutical drug and an effective amount of hyaluronic acid produced by hyaluronan synthase.
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48. A purified and isolated nucleic acid sequence encoding enzymatically active hyaluronan synthase, the nucleic acid sequence selected from the group consisting of:
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(a) the nucleic acid sequence in accordance with SEQ ID NO;
1;
(b) complementary nucleic acid sequences to the nucleic acid sequence in accordance with SEQ ID NO;
1;
(c) nucleic acid sequences which will hybridize to the nucleic acid in accordance with SEQ ID NO;
1;
(d) nucleic acid sequences which will hybridize to the complementary nucleic acid sequences of SEQ ID NO;
1; and
(e) nucleic acid sequences which will hybridize to PCR probes selected from the group consisting of PCR probes of SEQ ID NO;
3, SEQ ID NO;
4, SEQ ID NO;
5, SEQ ID NO;
6.
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49. A purified and isolated nucleic acid segment consisting essentially of a nucleic acid segment encoding enzymatically active hyaluronan synthase.
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51. An isolated nucleic acid segment consisting essentially of a nucleic acid segment encoding hyaluronan synthase having a nucleic acid segment sufficiently duplicative of the nucleic acid segment in accordance of SEQ ID NO:
- 1 to allow possession of the biological property of encoding for Streptococcus equisimilis hyaluronan synthase.
- View Dependent Claims (52, 53)
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54. A purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase, wherein the purified nucleic acid segment is capable of hybridizing to the nucleotide sequence in accordance with SEQ ID NO:
- 1.
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55. A purified nucleic acid segment according to SEQ ID NO:
- 3 capable of hybridizing to SEQ ID NO;
1.
- 3 capable of hybridizing to SEQ ID NO;
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56. A purified nucleic acid segment according to SEQ ID NO:
- 4 capable of hybridizing to SEQ ID NO;
1.
- 4 capable of hybridizing to SEQ ID NO;
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57. A purified nucleic acid segment according to SEQ ID NO:
- 5 capable of hybridizing to SEQ ID NO;
1.
- 5 capable of hybridizing to SEQ ID NO;
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58. A purified nucleic acid segment according to SEQ ID NO:
- 6 capable of hybridizing to SEQ ID NO;
1.
- 6 capable of hybridizing to SEQ ID NO;
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59. A purified nucleic acid segment having a coding region encoding enzymatically active hyaluronate synthase, the purified nucleic acid segment selected from the group consisting of:
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(A) the nucleic acid segment according to SEQ ID NO;
2;
(B) the nucleotide sequence in accordance with SEQ ID NO;
1;
(C) nucleic acid segments which hybridize to the nucleic acid segments defined in (A) or (B) or fragments thereof; and
(D) nucleic acid segments which but for the degeneracy of the genetic code, or encoding of functionally equivalent amino acids, would hybridize to the nucleic acid segments defined in (A), (B), and (C).
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Specification