ytgP

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

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C435S252300, C435S320100, C435S325000, C536S023700

Reexamination Certificate

active

06326172

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 ytgP (membrane protein) family, as well as their variants, herein referred to as “ytgP,” “ytgP polynucleotide(s),” and “ytgP polypeptide(s)” as the case may be.
BACKGROUND OF THE INVENTION
The Streptococci make up a medically important genera of microbes known to cause several types of disease in humans, including, for example, otitis media, conjunctivitis, pneumonia, bacteremia, menigitis, sinusitis, pleural empyema and endocarditis, and most particularly meningitidis, such as for example infection of cerebrospinal fluid. Since its isolation more than 100 years ago,
Streptococcus pneumoniae
has been one of the more intensively studied microbes. For example, much of our early understanding that DNA is, in fact, the genetic material was predicated on the work of Griffith and of Avery, Macleod and McCarty using this microbe. Despite the vast amount of research with
S. pneumoniae,
many questions concerning the virulence of this microbe remain. It is particularly preferred to employ Streptococcal genes and gene products as targets for the development of antibiotics.
The frequency of
Streptococcus pnezumoniae
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
Streptococcus pneumoniae
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 genonics 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 ytgP 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 ytgP, in particular ytgP polypeptides and ytgP 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 ytgP 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 ytgP polypeptides and polynucleotides as described in greater detail below. In particular, the invention relates to polypeptides and polynucleotides of a ytgP of
Streptococcus pneumoniae,
that is related by amino acid sequence homology to
B. subtilis
ytgP polypeptide. The invention relates especially to ytgP 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
ytgP Polynucleotide and Polypeptide Sequences
(A)
Streptococcus pneumoniae
ytgP polynucleotide sequence
[SEQ ID NO:1]
5′-
ATGTCGCACGAAAACAATCACCAGCAGGCCCAGATGTTACGGGGGACTGCTTGGCTAACGGCTAGTAACTTT

ATCAGTCGCCTACTCGGGGCTGTTTATATTATCCCTTGGTACATCTGGATGGGGGCTTATGCAGCTAAGGCA

AATGGTCTCTTTACCATGGGTTACAATATCTATGCTTGGTTCTTGTTGGTTTCAACAGCGGGGATTCCAGTT

GCGGTGGCCAAGCAAGTTGCCAAG

TATAATACCATGCGAGAAGAAGAGCATAGCTTTGCCCTGATTCGGAGCTTCTTAGGCTTTATGACAGGACTA

GGCCTGGTTTTTGCTTTAGTCTTGTATGTCTTTGCTCCTTGGCTAGCAGACTTGTCTGGCGTGGGCAAAGAC

TTGATCCCAATCATGCAAAGCTTGGCTTGGGGAGTCTTGATTTTCCCGTCTATGAGTGTTATCCGAGGATTT

TTCCAAGGGATGAATAACCTCAAA

CCCTATGCCATGAGCCAAATTGCTGAGCAGGTCATTCGTGTTATCTGGATGCTCCTAGCAACCTTTATCATT

ATGAAGCTCGGTTCAGGAGATTATCTAGCAGCCGTTACCCAATCAACCTTTGCTGCCTTTGTCGGTATGGTA

GCCAGTTTTGCAGTCTTGATTTATTTCCTTGCCCAAGAAGGTTCACTCAAAAGAGTCTTTGAAACAGGAGAT

AAGATTAACAGTAAGCGTCTCTTG

GTTGATACCACTAAGGAAGCCATTCCTTTTATCCTGACAGGGTCTGCCATCCAGCTCTTCCAGATTTTGGAT

CAGCTGACCTTTATCAATAGTATGAGCTGGTTTACCAACTACAGCAATGAGGACTTGGTTGTCATGTTTTCT

TATTTCTCAGCCAATCCTAATAAAATCACGATGATTTTGATTTCTGTAGGGGTTTCGATTGGGAGTGTTGGT

TTGCCACTTTTGACGGAAAACTAT

GTCAAGGGGGACTTGAAAGCAGCTTCTCGTCTCGTTCAGGACAGTCTCACCCTACTCTTTATGTTCTTGCTA

CCAGCAACGGTTGGAGTGGTTATGGTAGGAGAACCTCTTTATACGGTCTTCTATGGTAAGCCAGATAGTTTG

GCTCTGGGCTTATTTGTCTTTGCAGTTTTGCAGTCTATTATTTTAGGCTTGTACATGGTCTTGTCTCCAATG

CTTCAGGCCATGTTCCGCAACCGC

AAGGCCGTTCTCTATTTTATCTATGGTTCTATTGCCAAGCTAGTCTTGCAACTACCTACCATCGCCCTCTTC

CACAGTTATGGTCCTTTGATTTCAACAACCATTGCTCTCATCATTCCTAACGTCTTGATGTATCGGGATATT

TGTAAAGTAACTGGTGTCAAGCGCAAGGTGATTTTGAAGCGAACCATTTTAATCAGTTTGCTGACCCTAGTC

ATGTTTCTGTTAATAGGAACCATC

CAGTGGCTGTTAGGATTTTTCTTCCAACCAAGTGGACGTTTGTGGAGCTTCTTTTATGTAGCTCTTGTCGGT

GCCATGGGGGGTGGACTTTATATGGTTATGAGTCTGCGTACCTATTTATTAGATAAGGTAATAGGAAAAGCC

CAAGCAGATCGCCTGCGAGCAAAATTTAAGCTTTCGTAA-3′

(B)
Streptococcus pneumoniae
ytgP polypeptide sequence deduced from a
polynucleotide sequence in this table [SEQ ID NO:2].
NH
2
-
MSHENNHQQAQMLRGTAWLTASNFISRLLGAVYIIPWYIWMGAYAAKANGLFTMGYNIYAWFLLVSTAGIFV

AVAKQVAKYNTMREEEHSFALIRSFLGFMTGLGLVFALVLYVFAPWLADLSGVGKDLIFIMQSLAWGVLIFF

SMSVIRGFFQGMNNLKFYAMSQIAEQVIRVIWMLLATFIIMKLGSGDYLAAVTQSTFAAFVGMVASFAVLIY

FLAQEGSLKRVFETGDKINSKRLL

VDTTKEAIPFILTGSAIQLFQILDQLTFINSMSWFTNYSNEDLVVMFSYFSANFNKITMILISVGVSIGSVG

LFLLTENYVKGDLKAASRLVQDSLTLLFMFLLFATVGVVMVGEFLYTVFYGKFDSLALGLFVFAVLQSIILG

LYMVLSFMLQAMFRNRKAVLYFIYGSIAKLVLQLFTIALFHSYGFLISTTIALIIFNVLMYRDICKVTGVKR

KVILKRTILISLLTLVMFLLIGTI

QWLLGFFFQFSGRLWSFFYVALVGAMGGGLYMVMSLRTYLLDKVIGKAQADRLRAKFKLS-COOH
Deposited Materials
A deposit comprising a
Streptococcus pneumoniae
0100993 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 Apr. 11, 1996 and assigned deposit number 40794. The deposit was described as
Streptococcus pneumoniae
0100993 on deposit.
On Apr. 17, 1996 a
Streptococcus pneumoniae
0100993 DNA library in
E. coli
was similarly deposited with the NCIMB and assigned deposit number 40800. The
Streptococcus pneumoniae
strain deposit is referred t

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