WO1991009140A1 - Sonde de detection du syndrome de l'x fragile - Google Patents

Sonde de detection du syndrome de l'x fragile Download PDF

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Publication number
WO1991009140A1
WO1991009140A1 PCT/GB1990/001940 GB9001940W WO9109140A1 WO 1991009140 A1 WO1991009140 A1 WO 1991009140A1 GB 9001940 W GB9001940 W GB 9001940W WO 9109140 A1 WO9109140 A1 WO 9109140A1
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WIPO (PCT)
Prior art keywords
sequence
nucleic acid
fragile
oligonucleotide
acid fragment
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PCT/GB1990/001940
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English (en)
Inventor
Ruth Nerida Mackinnon
Mark Charles Hirst
Kay Elizabeth Davies
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Medical Research Council
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Publication date
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Publication of WO1991009140A1 publication Critical patent/WO1991009140A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Definitions

  • the present invention relates to nucleic acid fragments and their use in the diagnosis of fragile X syndrome.
  • the fragile X (Martin-Bell) syndrome (McKusick cat no: 30955) is characterised by mental retardation, and physical anomalies including large ears, oblong face and macro- orchidism in affected males.
  • lymphocytes derived from patients are cultured in thymidine deficient media, 2 to 50% of the cells express a fragile site at Xq27.3.
  • the locus DXS269 shows no recombination with the fragile site (Suthers et al. , Am. J. Hum. Genet.. in press 1989) but is not very informative and shows no differences in fragile X patients compared to normal individuals in conventional Southern blots or by pulsed field gel electrophoresis. These loci are being used clinically in conjunction with cytogenetic evaluation of the expression of the fragile site.
  • M54 which lies in the region of the fragile site closer than DXS304.
  • the locus is different from those previously reported for markers in this region such as DXS304, DXS296 and DXS396.
  • In situ hybridisation of radio-labelled M54 to a human male karyotype reveals grains in photographic emulsion at Xq27 and 12 autosomal sites, 9 of which are fragile sites and one of which is a telomere.
  • the present invention therefore provides an oligonucleotide or nucleic acid fragment comprising at least 17 contiguous nucleotide bases in a sequence:
  • Oligonucleotides and fragments according to the invention may be single or double stranded, RNA or DNA. When double stranded they contain at least 17 contiguous nucleotide base pairs. Preferred oligonucleotides and fragments of the invention are at least 20 bases or base pairs in length, more preferably at least 50 bases or base pairs in length and most preferably about 110 bases or base pairs in length although yet longer oligonucleotides and fragments are also within the scope of the invention.
  • oligonucleotides and fragments must contain at least 17 contiguous bases or base pairs having a sequence (a) , (b) or (c) above; oligonucleotides and fragments longer than 17 bases or base pairs preferably contain a sequence according to Figure 1, complementary thereto or hybridisable therewith of at least 20, more preferably at least 50 and most preferably about 110 bases or base pairs in length. Particularly preferred oligonucleotides and fragments of the invention contain the entire sequence of Figure 1 or are perfectly complementary thereto.
  • Oligonucleotides and fragments which contain a sequence hybridisable to a sequence of at least 17 contiguous nucleotide bases or base pairs of Figure 1, or a complementary sequence will hybridise at least under low stringency conditions defined for the present invention as 3 x SSC at 65°C. More preferably such oligonucleotides and fragments will hybridise under high stringency conditions, i.e, 0.1 x SSC at 65°C oligonucleotides and fragments hybridising under low stringency must be at least 70% homologous with the corresponding portion of the sequence of Figure 1 (or the complementary sequence) .
  • To hybridise at high stringency it is necessary for there to be 90 or 95% ho ology. Differences may arise by way of substitution, deletion or insertion.
  • the oligonucleotides fragments of the invention are capable of hybridising selectively with the human X chromosome in the region Xq27.3.
  • the oligonucleotides and fragments of the invention hybridise with the Xq27.3 region under low or high stringency conditions. Under low stringency conditions hybridisation with chromosomal DNA blots will occur between the oligonucleotides or fragments of the invention and a large class of sites of related sequence in various parts of the human genome. Such hybridisation has uses in identifying fragile sites other than the fragile X site and mobile regions with the human genome. Under increasingly high stringency conditions the cross-hybridisation is progressively reduced and at the highest stringency (0.1 x SSC at 65°C) only a few sites are identified.
  • Hybridisation at high stringency reveals a number of loci related to M54 adjacent the fragile X site with Xq27.3.
  • Particular oligonucleotides and fragments of the invention therefore include several copies of the sequence of Figure 1, or complementary sequences or sequences hybridisable thereto. Two of these sites are located either side of the fragile X site itself and thus define a region containing the fragile X site. Fragments comprising the whole of the region between these two sites and optionally also containing flanking DNA from one or both of the proximal and distal regions are particularly useful as probes for the fragile X site and because such fragments can be used to identify coding sequences disrupted by the fragile X site.
  • oligonucleotides and fragments of the invention are double stranded DNA fragments containing the full sequence of Figure 1 and capable of hybridising under high stringency conditions at a M54 locus.
  • oligonucleotides and fragments of the invention comprise the sequence (I) :
  • X is A or C (using the internationally recognised 1- letter codes for nucleotide bases) or the sequence (II) :
  • Y is T or G.
  • Sequence (I) appears twice in the M54 locus sequence of Figure 1 and is believed to be unique. Oligonucleotides and fragments containing this sequence are therefore useful as probes for the M54 locus. Preferred such oligonucleotides and fragments contain two copies of the sequence (I) , more preferably spaced apart by about 50 nucleotide residues, most preferably spaced apart by the 50 residues in the sequence of Figure 1. Sequence (II) is complementary to sequence (I) and oligonucleotides and fragments containing this sequence, or preferred such oligonucleotides and fragments as described for sequence (I) are also useful as probes.
  • the oligonucleotides and fragments of the invention For use as probes, it will be necessary for the oligonucleotides and fragments of the invention to be detectable following hybridisation with human DNA and this may be achieved by any known labelling technique.
  • the probe will be labelled by incorporation of radio- isotopes such as 32 P, for instance by nick translation, which may be detected by auto-radiography.
  • Other labelling techniques include provision of directly detectable labels such as fluorescent labels and indirectly detectable labels such as biotin or enzyme labels. A skilled person will be aware of the techniques required for introduction of these and other known labels and for the detection of such labels.
  • the invention therefore also provides a probe comprising an oligonucleotide or nucleic acid fragment as defined above having a detectable label attached thereto.
  • the M54 locus comprises a region of the X chromosome extending around the fragile X site and including the sequence of Figure 1. It does not overlap with any previously reported locus. Digests of the human X chromosome obtained using the restriction enzymes Rsa I, Pvu II, ECoRI or Hind III produced by conventional techniques yield a variety of DNA fragments, each of which hybridises with the M54 locus.
  • the M54 locus is, therefore, defined as that portion of the Xq27-qter region of the human X chromosome where one or more of the aforementioned Rsa I, Pvu II, EcoRI or Hind III fragments hybridise.
  • the present invention further provides oligonucleotide and nucleic acid fragments and probes which hybridise with the M54 locus as defined above and/or with any one or more of the aforementioned Rsa I, Pvu II, EcoRI and Hind III fragments.
  • the aforementioned Rsa I, Pvu II, EcoRI or Hind III fragments are all examples of fragments according to the present invention. Using known techniques, these fragments and probes may be employed to identify further fragments which will hybridise to the human X chromosome between the fragile X site and the M54 locus as defined.
  • Oligonucleotides and fragments and probes which hybridise to the human X chromosome between the fragile X site and the M54 locus as defined form a further aspect of the invention. Some such oligonucleotides and fragments will hybridise with one or more of the aforementioned Rsa I, Pvu II, EcoRI and Hind III fragments. Others of the oligonucleotides and fragments which hybridise between the fragile X site and the M54 locus will not hybridise to any of the aforementioned fragments but will nevertheless be useful in probing human X chromosomes for diagnosis of fragile X disease because they are capable of identifying regions nearer the fragile X site than the M54 locus.
  • oligonucleotides and fragments according to the present invention which hybridise at the M54 locus or between the fragile X site and the M54 locus may also find uses in diagnosis of other X-linked disorders.
  • use of oligonucleotides and fragments according to the invention to detect other fragile sites and mobile elements elsewhere in the human genome may enable diagnosis and therapy of various other diseases such as cancers and other syndromes which arise from rearrangements involving human chromosome 2.
  • Oligonucleotides and fragments containing a mutant of the Fig.l sequence and oligonucleotides or fragments of the invention containing to Fig.l sequence or a part thereof and mutant flanking sequences are therefore also useful in diagnosis or therapy of X-linked disorders in accordance with the invention.
  • Hybridisation as referred to herein is conducted by admixing the oligonucleotides or fragments to be hybridised at 65°C in a buffer comprising SSC (at a specified concentration), 0.1% sodium dodecyl sulphate (SDS), 10% dextran sulphate and 1 x Denhardts solution followed by washing in high or low stringency buffer at 65 ⁇ C.
  • 1 x SSC is 0.15M sodium chloride, 0.015. sodium citrate, pH 7.0 buffer solution.
  • Denhardts solution is 0.02% polyvinyl pyrrolidine, 0.02% Ficoll 400 and 0.2% bovine serum albumin (BSA) .
  • Low stringency buffer comprises 3 x SSC and 0.1% SDS whereas high stringency buffer comprises 0.1 x SSC and 0.1% SDS.
  • .•Oligonucleotides and fragments according to the invention may be obtained by digestion of human chromosome X or, preferably of the Xq27-qter region of the X chromosome, cloning the oligonucleotides and fragments as necessary and selecting oligonucleotides and fragments capable of hybridisation around Xq27.3, for instance at M54.
  • a skilled person would be well aware of the techniques required to conduct the digestion, cloning and screening to produce oligonucleotides and fragments as hereinbefore defined.
  • the oligonucleotide and nucleic acid fragments of the present invention particularly when bearing a detectable label, may be used for the diagnosis of fragile X syndrome.
  • the invention therefore further provides a process for diagnosing fragile X syndrome comprising contacting DNA from an individual potentially at risk of contracting fragile X syndrome with an oligonucleotide or nucleic acid fragment or probe as hereinbefore defined under suitable hybridising conditions. Hybridisation of the fragment or probe may be detected by conventional techniques.
  • the diagnostic process of the invention preferably further comprises similar screening of DNA samples from close and distant relatives of the individual potentially at risk, particularly grandparents, parents, aunts and uncles, siblings and/or children of the individual.
  • the DNA samples used may be freshly obtained or the process may be applied to archival material such as that obtained shortly after birth as'part of the Guthrie test.
  • Such testing of relatives, coupled with case histories indicating the emergence of fragile X syndrome is used to trace the susceptible DNA through the family tree based on patterns of hybridisation to individual copies of M54 or related sequences in the vicinity of the fragile X site and thus determine whether the individual potentially at risk has, or has not inherited a susceptible X chromosome.
  • the diagnostic process is conducted to screen for carriers of fragile X syndrome and/or as a pre-natal screening to identify at-risk foetuses.
  • Oligonucleotides and fragments and probes according to the present invention may also be used in similar manner to diagnose other X-linked disorders as mentioned above.
  • the invention further provides a kit for conducting a diagnostic test for fragile X syndrome comprising an oligonucleotide or nucleic acid fragment or probe as hereinbefore defined and one or more of the following accessory components: A reagent or reagents for labelling the oligonucleotide or nucleic acid fragment; one or more restriction enzymes for the digestion of the patient's DNA; a buffer or buffers for conducting the digestion and/or loading the digestion DNA onto a support or substrate for hybridisation; a support or substrate such as a hybridisation filter on which to conduct hybridisation; a buffer or buffers for washing the hybridisation filter; a buffer or buffers for conducting hybridisation under low or high stringency conditions; a reagent or reagents for detecting a probe as hereinbefore defined; and control reagents such as fragile X positive and/or negative samples of human DNA and/or samples of non-human DNA, as well as standards for comparison of the signal form a detectable label.
  • the M54 locus of the human X chromosome has been found to have a highly conserved sequence (it also appears in mouse, pig, cow, baboon and chicken and yeast chromosomes) indicating that the M54 locus may be functionally important and be adjacent to or part of a gene sequence. It is reasonable to predict that the gene or genes at or adjacent the M54 locus encode one or more proteins and that these proteins will be capable of raising antibodies when used as i munogens.
  • the genes may be obtained and expressed by conventional techniques and proteins expressed by the genes may be used to raise antibodies by well known methods.
  • the proteins and antibodies against the proteins may, themselves, have uses in the diagnosis of fragile X syndrome.
  • the present invention provides: cloning and expression vectors containing one or more oligonucleotide or nucleic acid fragment of the present invention as hereinbefore defined, such as a gene corresponding to the M54 locus or cDNA corresponding to such a gene, the expression vector further comprising initiation and termination signals, regulatory and promoter sequences in correct position, orientation and reading frame as appropriate; host cells transformed with said cloning or expression vectors; polypeptides or proteins expressed by culturing such transformed host cells containing an expression vector; polyclonal and monoclonal antibodies against such polypeptides and proteins and antibody producing cells, such as hybridomas, capable of secreting such antibodies.
  • the complementary strand nucleotide base sequence of Fig. 1 contains two open reading frames and the sequence encodes polypeptides of amino acid residue sequences shown in Fig. 3. Such polypeptides are useful in generating antibodies which can be used in research and may also be used diagnostically or therapeutically.
  • the present invention therefore provides a polypeptide or protein comprising at least 5 contiguous amino acid residues in a sequence according to Fig. 3.
  • the polypeptide or protein comprises at least 10 contiguous amino acid residues in a sequence according to Fig. 3 more preferably at least 20, for example at least 30 and most preferably all of the amino acid residues shown in Fig. 3.
  • Polypeptides of the invention longer than 5 amino acid residues may include portions homologous to further portions of the sequence of Fig. 3, differing from that sequence by substitutions, deletions and/or insertions provided that they contain a contiguous sequence of 5 or more amino acid residues identical to the sequence of Fig. 3.
  • the invention further provides antibodies, whether polyclonal, monoclonal or produced by recombinant DNA techniques, against proteins or polypeptides as defined above.
  • FIG 1. shows the sequence of the M54 locus (one strand only) and, boxed, sequences (I) appearing therein.
  • FIG 2. shows the in situ hybridisation pattern produced according to Example 1 below.
  • FIG 3. shows the amino acid residue sequences encoded by the various reading frames in the nucleic acid strand of Fig. 1.
  • Labelled probe was lyophilized, resuspended at 0.3 ⁇ g/ml in hybridisation buffer containing 5Q% formamide, 5 x Dehardt's solution, 5 x SSPE (0.9M NaCl/50mM NaH 2 P0 4 /5 mM EDTA), pH 7.2, 10% dextran sulfate, an 200 ug/ml salmon solution was placed on each slide, covered with a cover ⁇ lip, and incubated overnight at 43°C. Subsequently, the slides were rinsed in 5 x SSC to remove the coverslips and washed in three changes of 2 x SSC at room temperature over a period of 2h. The slides were then washed in 1 x SSC at 65°C for lh, followed by 0.2 x SSC and 0.1 x SSC for 30 min (each at room temperature) and finally dehydrated.
  • hybridisation buffer containing 5Q% formamide, 5 x Dehardt's solution, 5 x SSPE (
  • the slides were then dipped in Ilford L4 emulsion, exposed for 14 days at 4°C, developed in Kodak D19 for 7 min at 20°C, stained with Hoechst 33258 (10 ⁇ g/ l in 2 x SSC) for 30 min, exposed to UV light in 2 X SSC for lh, and then stained with 10% Giemsa for 15 min.
  • the resulting silver grain distribution pattern is shown in Fig. 2. Only grains actually touching the chromosomes were scored. Clusters of grains were noted, but scored as a single hybridisation event on the ideograms illustrating grain distribution. This procedure enabled the ready identification of G-banded chromosomes and the distribution of silver grains on them.
  • Fig. 2 Only grains actually touching the chromosomes were scored. Clusters of grains were noted, but scored as a single hybridisation event on the ideograms illustrating grain distribution. This procedure enabled the ready identification of G-banded chromosomes and the distribution of silver grains on them.
  • FIG. 2 shows the pattern of grains in an autoradiograph of the spread. Grain distribution in 53 cells following jLn situ hybridisation of M54 to replication-banded normal male chromosomes. Closed triangles _i denote sites of hybridisation which are equal to or greater than that scored Xq2 ' 7, the location of the FRAXA locus. Open triangles A denote where these sites of hybridisation corresponds to the location of a fragile site.

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Abstract

Oligonucléotide ou fragment d'acide nucléique comprenant au moins 17 bases de nucléotides contiguës et organisé en séquence: (a) correspondant à au moins 17 bases de nucléotides de la séquence (α), (b) complémentaire d'une séquence (a), ou (c) hybridable avec une séquence (a) ou (b). La séquence (a) ou (b) est utile dans le diagnostic du syndrome de l'X fragile.
PCT/GB1990/001940 1989-12-12 1990-12-12 Sonde de detection du syndrome de l'x fragile WO1991009140A1 (fr)

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GB8928029.1 1989-12-12
GB898928029A GB8928029D0 (en) 1989-12-12 1989-12-12 Probe

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2672618A1 (fr) * 1991-02-13 1992-08-14 Centre Nat Rech Scient Fragment d'acide nucleique de la region du chromosome x implique dans le syndrome x fragile, sonde nucleotidique et procede pour le diagnostic du retard mental avec x fragile.
WO1992014840A1 (fr) * 1991-02-13 1992-09-03 Institut National De La Sante Et De La Recherche Medicale Fragment d'acide nucleique de la region du chromosome x implique dans le syndrome x fragile, sonde nucleotidique et procede pour le diagnostic du retard mental avec x fragile
EP0724646A4 (fr) * 1991-01-04 1995-06-09 Univ Washington Sequences d'adn associees au syndrome de chromosome x fragile isole
US5698686A (en) * 1994-10-20 1997-12-16 Arch Development Corporation Yeast telomerase compositions
US5741645A (en) * 1993-06-29 1998-04-21 Regents Of The University Of Minnesota Gene sequence for spinocerebellar ataxia type 1 and method for diagnosis
US5876949A (en) * 1995-05-31 1999-03-02 The Trustees Of The University Of Pennsylvania Antibodies specific for fragile X related proteins and method of using the same

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1986005512A1 (fr) * 1985-03-13 1986-09-25 Centre National De La Recherche Scientifique (Cnrs Sonde adn destinee au diagnostic antenatal de certaines anomalies chromosomiques
WO1988003572A1 (fr) * 1986-11-13 1988-05-19 Kay Elizabeth Davies Sequence d'adn
WO1990005194A1 (fr) * 1988-11-01 1990-05-17 Medical Research Council Sonde permettant de detecter la presence d'un syndrome x fragile

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1986005512A1 (fr) * 1985-03-13 1986-09-25 Centre National De La Recherche Scientifique (Cnrs Sonde adn destinee au diagnostic antenatal de certaines anomalies chromosomiques
WO1988003572A1 (fr) * 1986-11-13 1988-05-19 Kay Elizabeth Davies Sequence d'adn
WO1990005194A1 (fr) * 1988-11-01 1990-05-17 Medical Research Council Sonde permettant de detecter la presence d'un syndrome x fragile

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Genomics, volume 4, April 1989, Academic Press, Inc., M.N. Patterson et al.: "Genetic and physical mapping of a novel region close to the fragile X site on the human X chromosome", pages 570-578 *
Science, volume 246, 8 December 1989, G.K. Suthers et al.: "A new DNA marker tightly linked to the fragile X locus (Fraxa)", pages 1298-1300 *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0724646A4 (fr) * 1991-01-04 1995-06-09 Univ Washington Sequences d'adn associees au syndrome de chromosome x fragile isole
EP0724646A1 (fr) * 1991-01-04 1996-08-07 Washington University Sequences d'adn associees au syndrome de chromosome x fragile isole
US6197500B1 (en) 1991-01-04 2001-03-06 Adelaide Medical Centre For Women And Children DNA sequences related to fragile X syndrome
US6242576B1 (en) 1991-01-04 2001-06-05 Women's And Children's Hospital Monoclonal and polyclonal antibodies relating to fragile X syndrome
FR2672618A1 (fr) * 1991-02-13 1992-08-14 Centre Nat Rech Scient Fragment d'acide nucleique de la region du chromosome x implique dans le syndrome x fragile, sonde nucleotidique et procede pour le diagnostic du retard mental avec x fragile.
WO1992014840A1 (fr) * 1991-02-13 1992-09-03 Institut National De La Sante Et De La Recherche Medicale Fragment d'acide nucleique de la region du chromosome x implique dans le syndrome x fragile, sonde nucleotidique et procede pour le diagnostic du retard mental avec x fragile
US5741645A (en) * 1993-06-29 1998-04-21 Regents Of The University Of Minnesota Gene sequence for spinocerebellar ataxia type 1 and method for diagnosis
US5698686A (en) * 1994-10-20 1997-12-16 Arch Development Corporation Yeast telomerase compositions
US5916752A (en) * 1994-10-20 1999-06-29 Arch Development Corporation Telomerase screening methods
US6387619B1 (en) 1994-10-20 2002-05-14 Arch Development Telomerase compositions and methods
US5876949A (en) * 1995-05-31 1999-03-02 The Trustees Of The University Of Pennsylvania Antibodies specific for fragile X related proteins and method of using the same

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GB8928029D0 (en) 1990-02-14

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