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
1999-01-26
2002-09-10
Duffy, Patricia A. (Department: 1645)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
C435S069300, C435S070100, C435S071100, C435S320100, C435S325000, C435S254110, C435S252300, C536S023700
Reexamination Certificate
active
06448038
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 polypeptides of the birA (biotin ligase/biotin holoenzyme synthetase) family, as well as their variants, herein referred to as “birA,” “birA polynucleotide(s),” and “birA 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 skin 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 birA 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 birA, in particular birA polypeptides and birA 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 birA expression or activity.
Various changes and modifications within 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 birA polypeptides and polynucleotides as described in greater detail below. In particular, the invention relates to polypeptides and polynucleotides of a birA of
Staphylococcus aureus
, which is related by amino acid sequence homology to birA of
Bacillus subtilis
polypeptide. The invention relates especially to birA 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
birA Polynucleotide and Polypeptide Sequences
(A)
Staphylococcus aureus
birA polynucleotide sequence [SEQ ID NO: 1].
5′-
ATGTCAAAATATAGTCAAGATGTACTTCAATTACTCTATAAAAATAAACCGAATTATATATCTGGACAAAGC
ATTGCGGA
ATCACTTAATATtTCaCGCACTGCAGTAAAAAAgGTGATTGACCAATTAAAGTTAGAGGGATGTAAAATtGA
TTCAGTAA
ATCATAAAGGgCATTTATTACAACAaCTCCCAGATaTTTGGTATCAgGGTATAATAGACCAATATACAAAAA
GTTCTGCT
TTGTTTGATTTTAGTGAAGTaTACGATTCAATaGATTCTACACAACTTGCTGCGAAAAAGTCACTTGTTGGA
AATCAATC
TTCATTTTTTATCTTGAGTGATGAACAAACGAAAGGTCGTGGGCGATTTAATAGACATTGGAGTTCTTCAAA
AGGGCAAG
GACTTTGGATGTCtGTCGTgTTAAGACCTAACGTTGCATTCTCAATGATATCTAAATTTAATTTATTTATTG
CATTAGGG
ATAAGAGATGCGATTCAACATTTTAGTCAAGATGAaGTCAAAGTGAAATGGCCGAATGATATATATATTGAT
AATGGTAA
AGTGTGTGGTTTCTTAACTGAAATGGTTGCTAATAATGATGGTATAGAAGCAATAATATGTGGTATAGGTAT
TAATTTGA
CGCAACAACTAGAAaACTTTGATGAAAGTATTAGACATAGAGCAACAAGTATACAATTACATGATAAAAATA
AATTAGAT
AGATATCAATTTTTAGAGAGATTACTTCAAGAAATTGAAAAAAGATATAATCAATTTTTAACGTTACCTTTT
TCTGAAAT
TCGTGAAGAATATATTGCAGCTTCTAATATTTGGAATAGAACGTTGCTATTTACAGAAAATGATAAACAGTT
TAAAGGAC
AAGCAATTGATTTAGATTACGATGGCTATCTAATTGTTAGAGATGAAGCGGGTGAATCACACCGTTTAATTA
GTGCAGAT
ATAGATTTTTAA-3′
(B)
Staphylococcus aureus
birA polypeptide sequence deduced
from a polynucleotide sequence in this table [SEQ ID NO:2].
NH
2
MSKYSQDVLQLLYKNKPNYISGQSIAESLNISRTAvKKVIDQLKLEGCKIBSVNHKGHLL
QQLPDIWYQGIIDQYTKSSALFDFSEVYDSIDSTQLAAKKSLVGNQSSFFILSDEQTKGR
GRFNRHWSSSKGQGLWMSWLRPNVAFSMISKFNLFIALGIRDAIQHFSQDEVKVKWPND
IYIDNGKVCGFLTEMVANNDGIEAIICGIGINLTQQLENFDESIRHRATSIQLHDKNKLD
RYQFLERLLQEIEKRYNQFLTLPFSEIREEYIAASNIWNRTLLFTENDKQFKGQAIDLDY
DGYLIVRDEAGESHRLISADIDF-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 birA 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 on the International Recognition of the Deposit of Micro-organisms for Purposes of Patent Procedure. The deposited strain still be irrevocably and without restriction or condition released to the public upon the issuance of a patent. The deposited strain is provided merely as convenience to those of skill in the art and is not an admission that a deposit is required for enablement, such as that required under 35 U.S.C. §112. A license may be required to make, use or sell the deposited strain, and compounds derived therefrom, and no such license is hereby granted.
In one aspect of the invention there is provided an isolated nucle
Horn Stephanie Van
Huang Jianzhong
Kallender Howard
McDevitt Damien
Sylvester Daniel R
Duffy Patricia A.
Fedon Jason C.
Gimmi Edward R.
Kinzig Charles M.
SmithKline Beecham Corporation
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