Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Reexamination Certificate
1999-06-29
2001-06-26
Priebe, Scott D. (Department: 1632)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
C435S252300, C435S325000, C435S320100, C435S455000, C435S471000, C536S023100, C536S023400, C536S023700
Reexamination Certificate
active
06251633
ABSTRACT:
FIELD OF THE INVENTION
This invention relates to newly identified polynucleotides and polypeptides, and their production and uses, as well as their variants, agonists and antagonists, and their uses. In particular, the invention relates to polynucleotides and poly,peptides of the Cell division family, as well as their variants, herein referred to as “FtsA,” “FtsA polynucleotide(s),” and “FtsA polypeptide(s)” as the case may be.
BACKGROUND OF THE INVENTION
It is particularly preferred to employ Staphylococcal genes and gene products as targets for the development of antibiotics. The Staphylococci make up a medically important genera of microbes. They, are known to produce two types of disease, invasive and toxigenic. Invasive infections are characterized generally by abscess formation effecting both ski surfaces and deep tissues. S. aureus is the second leading cause of bacteremia in cancer patients. Osteomyelitis, septic arthritis, septic thrombophlebitis and acute bacterial endocarditis are also relatively common. There are at least three clinical conditions resulting from the toxigenic properties of Staphylococci. The manifestation of these diseases result from the actions of exotoxins as opposed to tissue invasion and bacteremia. These conditions include: Staphylococcal food poisoning, scalded skin syndrome and toxic shock syndrome.
The frequency of
Staphylococcus aureus
infections has risen dramatically in the past few decades. This has been attributed to the emergence of multiply antibiotic resistant strains and an increasing population of people with weakened immune systems. It is no longer uncommon to isolate
Staphylococcus aureus
strains that are resistant to some or all of the standard antibiotics. This phenomenon has created an unmet medical need and demand for new anti-microbial agents, vaccines, drug screening methods, and diagnostic tests for this organism.
Moreover, the drug discovery process is currently undergoing a fundamental revolution as it embraces “functional genomics,” that is, high throughput genome- or gene-based biology. This approach is rapidly superseding earlier approaches based on “positional cloning” and other methods. Functional genomics relies heavily on the various tools of bioinformatics to identify gene sequences of potential interest from the many molecular biology databases now available as well as from other sources. There is a continuing and significant need to identify and characterize further genes and other polynucleotides sequences and their related polypeptides, as targets for drug discovery.
Clearly, there exists a need for polynucleotides and polypeptides, such as the FtsA embodiments of the invention, that have a present benefit of, among other things, being useful to screen compounds for antimicrobial activity. Such factors are also useful to determine their role in pathogenesis of infection, dysfunction and disease. There is also a need for identification and characterization of such factors and their antagonists and agonists to find ways to prevent, ameliorate or correct such infection, dysfunction and disease.
SUMMARY OF THE INVENTION
The present invention relates to FtsA, in particular FtsA polypeptides and FtsA polynucleotides, recombinant materials and methods for their production. In another aspect, the invention relates to methods for using such polypeptides and polynucleotides, including treatment of microbial diseases, amongst others. In a further aspect, the invention relates to methods for identifying agonists and antagonists using the materials provided by the invention, and for treating microbial infections and conditions associated with such infections with the identified agonist or antagonist compounds. In a still further aspect, the invention relates to diagnostic assays for detecting diseases associated with microbial infections and conditions associated with such infections, such as assays for detecting FtsA expression or activity.
Various changes and modifications with the spirit and scope of the disclosed invention will become readily apparent to those skilled in the art from reading the following descriptions and from reading the other parts of the present disclosure.
DESCRIPTION OF THE INVENTION
The invention relates to FtsA polypeptides and polynucleotides as described in greater detail below. In particular, the invention relates to polypeptides and polynucleotides of a FtsA of
Staphylococcus aureus,
that is related by amino acid sequence homology to actin/heat shock protein 70 polypeptide. The invention relates especially to FtsA having a nucleotide and amino acid sequences set out in Table 1 as SEQ ID NO:1 and SEQ ID NO:2 respectively. Note that sequences recited in the Sequence Listing below as “DNA” represent an exemplification of the invention, since those of ordinary skill will recognize that such sequences can be usefully employed in polynucleotides in general, including ribopolynucleotides.
TABLE 1
FtsA Polynucleotide and Polypeptide Sequences
(A)
Staphylococcus aureus
FtsA polynucleotide
sequence [SEQ ID NO: 1].
5′-
ATGGAAGAACATTACTACGTAAGTATTGATATTGGATCATCAAGCGTAA
AAACAATAGTAGGCGAGAAATTTCACAATGGTATAAATGTGATAGGTAC
AGGACAAACCTACACGAGCGGTATAAAAAATGGTTTAATTGATGATTTT
GATATTGCGCGACAAGCAATCAAAGACACAATTAAAAAGGCATCAATCG
CTTCGGGTGTTGATATTAAAGAAGTTTTCCTGAAATTACCTATC
ATTGGAACGGAAGTTTATGATGAATCAAATGAAATCGACTTTTATGAGG
ATACAGAAATCAACGGTTCACATATCGAAAAAGTATTAGAAGGTATTAG
AGAAAAAAATGATGTGCAAGAAACAGAAGTAATTAATGTGTTCCCGATT
CGTTTTATAGTCGATAAAGAAAATGAGGTTTCAGACCCTAAAGAATTAA
TTGCCAGACATTCATTAAAGGTTGAAGCAGGCGTAATTGCTATT
CAAAAATCGATTTTAATTAATATGATTAAATGCGTAGAAGCATGTGGTG
TTGATGTATTAGATGTTTACTCTGATGCATATAACTATGGTTCAATCCT
AACAGCTACTGAAAAAGAGTTAGGTGCATGTGTCATTGATATTGGTGAA
GACGTTACGCAAGTTGCTTTTTATGAACGCGGTGAATTAGTAGATGCTG
ATTCTATCGAAATGGCAGGGCGTGATATTACAGACGATATTGCA
CAAGGATTAAACACTTCTTATGAAACTGCTGAAAAAGTTAAACACCAAT
ATGGTCATGCATTCTATGATTCTGCTTCAGATCAAGATATCTTCACTGT
TGAACAGGTTGATAGTGATGAAACAGTACAGTATACTCAAAAAGATTTG
AGTGACTTTATTGAAGCGCGTGTAGAAGAAATATTCTTCGAAGTATTTG
ATGTTTTACAAGATTTAGGATTAACAAAAGTAAATGGTGGGTTT
ATTGTAACTGGTGGATCTACAAACTTACTTGGCGTAAAAGAATTATTAT
CAGATATGGTAAGTGAAAAAGTTAGAATTCACACGCCATCACAAATGGG
AATTAGAAAACCTGAATTTTCTTCAGCAATTTCTACAATTTCTAGTAGT
ATCGCTTTTGATGAGTTATTAGATTATGTTACAATTAATTATCATGATA
ATGAAGAAACTGAAGAAGATGTTATTGATGTGAAAGACAAAGAT
AACGAATCTAAATTAGGCGGATTTGATTGGTTTAAACGTAAAACAAACA
AAAAAGATACTCATGAAAATGAAGTAGAGTCAACAGATGAAGAAATTTA
TCAATCAGAAGATAATCATCAGGAACATAAACAGAATCATGAACATGTT
CAAGACAAAGATAAAGATAAAGAAGAAAGTAAATTCAAAAAACTAATGA
AATCTCTATTTGAATGA-3′
(B)
Staphylococcus aureus
FtsA polypeptide
sequence deduced from a polynucleotide sequence
in this table [SEQ ID NO:2].
NH
2
-
MEEHYYVSIDIGSSSVKTIVGEKFHNGINVIGTGQTYTSGIKNGLIDDF
DIARQAIKDTIKKASIASGVDIKEVFLKLPIIGTEVYDESNEIDFYEDT
EINGSHIEKVLEGIREKNDVQETEVINVFPIRFIVDKENEVSDPKELIA
RHSLKVEAGVIAIQKSILINMIKCVEACGVDVLDVYSDAYNYGSILTAT
EKELGACVIDIGEDVTQVAFYERGELVDADSIEMAGRDITDDIA
QGLNTSYETAEKVKHQYGHAFYDSASDQDIFTVEQVDSDETVQYTQKDL
SDFIEARVEEIFFEVFDVLQDLGLTKVNGGFIVTGGSTNLLGVKELLSD
MVSEKVRIHTPSQMGIRKPEFSSAISTISSSIAFDELLDYVTINYHDNE
ETEEDVIDVKDKDNESKLGGFDWFKRKTNKKDTHENEVESTDEEIYQSE
DNHQEHKQNHEHVQDKDKDKEESKFKKLMKSLFE-COOH
Deposited Materials
A deposit comprising a
Staphylococcus aureus
WCUH 29 strain has been deposited with the National Collections of Industrial and Marine Bacteria Ltd. (herein “NCIMB”), 23 St. Machar Drive, Aberdeen AB2 1RY, Scotland on Sep. 11, 1995 and assigned NCIMB Deposit No. 40771, and referred to as Staphylococcus aureus WCUH29 on deposit. The
Staphylococcus aureus
strain deposit is referred to herein as “the deposited strain” or as “the DNA of the deposited strain.”
The deposited strain comprises a full length FtsA gene. The sequence of the polynucleotides comprised in the deposited strain, as well as the amino acid sequence of any polypeptide encoded thereby, are controlling in the event of any conflict with any description of sequences herein.
The deposit of the deposited strain has been made under the terms of the Budapest Treaty o
Pearce, Jr. Kenneth H
Yan Kang
Deibert Thomas S.
Gimmi Edward R.
King William T.
Priebe Scott D.
SmithKline Beecham Corporation
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