Hyaluronate synthase gene and uses thereof
View Patent ↗Disclosed are DNA sequences encoding hyaluronic acid synthase that are employed to construct recombinant cells useful in the production of hyaluronate synthase and hyaluronic acid (HA). In preferred aspects, chromosomal DNA encoding the HA synthase gene, hasA, was cloned from a Streptococcus pyogenes genomic library. These vectors were used to transform host cells such as E. coli and acapsular Streptococci to produce hyaluronic acid. Resultant transformants were screened to identify colonies which have incorporated HA synthase DNA in a form that is being actively transcribed into the corresponding HA synthase enzyme. These colonies were selected and employed in the production of hyaluronic acid.
1. A method of detecting a DNA species comprising the steps of:
(a) obtaining a DNA sample;
(b) contacting said DNA sample with a nucleic acid segment in accordance with SEQ ID NO:1 under conditions effective to allow hybridization to form a complex; and
(c) detecting said complex.
2. A method for detecting a bacterial cell that expresses hyaluronic acid synthase, comprising the steps of:
(a) obtaining a bacterial cell sample suspected of expressing hyaluronan synthase;
(b) contacting nucleic acids from said cell sample with a nucleic acid segment in accordance with SEQ ID NO:1 under conditions effective to allow hybridization of substantially complementary nucleic acid sequences; and
(c) identifying the presence of hybridized complexes containing substantially complementary nucleic acid sequences within said sample; wherein the presence of increased levels of said substantially complementary nucleic acid sequence is indicative of a bacterial strain that expresses hyaluronan synthase.
3. The method of claim 1 , wherein the nucleic acids contacted are located within said cell.
4. The method of claim 1 , wherein the nucleic acids are separated from said cell prior to contact.
5. The method of claim 1 , wherein the nucleic acid segment comprises a detectable label and hybridized complexes are detected by detecting said label.
6. The method of claim 2 , wherein said nucleic acid segment comprises a radio-, an enzymatic-, a fluorescent-, a biotinyl-, or a chemiluminescent-label.