Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reissue Patent
2000-05-11
2003-07-01
Fredman, Jeffrey (Department: 1637)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C435S091100, C435S091200, C435S242000, C435S320100, C435S007210, C435S007100, C435S069100, C536S023100, C536S024300
Reissue Patent
active
RE038168
ABSTRACT:
BACKGROUND OF THE INVENTION
Dissection of germline mutations in a sensitive and specific manner presents a continuing challenge. In dominantly inherited diseases, mutations occur in only one allele and are often masked by the normal allele. For example, it is estimated that 20-40% of both APC and hMSH2 mutations are difficult or impossible to detect with standard techniques based on PCR analysis of genomic DNA or RNA transcripts
6-10
. Thus there is a need in the art for a technique which is relatively simple to perform and which will detect a broad spectrum of mutations in genes of clinical interest.
SUMMARY OF THE INVENTION
It is an object of the invention to provide a method for detecting mutations in the germline.
It is another object of the invention to provide a method for detecting mutations which are not detected by standard methods.
It is yet another object of the invention to provide a method for detecting mutations which lead to diminished or loss of expression of a gene product.
These and other objects of the invention are provided by one or more of the embodiments described below. In one embodiment a method of detecting mutations in a gene of interest on a chromosome of a human is provided. The method comprises the steps of:
obtaining cells of the human;
fusing said cells to rodent cell recipients to form a human-rodent cell hybrid;
testing said human-rodent cell hybrid to confirm the presence of said chromosome of the human in said hybrid;
testing said hybrid which contains said chromosome to detect a protein product of said gene, absence of said protein product or diminished amounts of said protein product indicating the presence of a mutation in the gene of interest of the human.
The present invention thus provides the art with a useful diagnostic tool in the evaluation of inherited diseases.
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Acta Universitatis Oulvensis SereiaLeila P(214) 4-54 1990.*
Mammalian GenomeBrown et al.5(7):434-437 (1994).*
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Nature Genetics,Ishioka et al.5: 124-129 (1993).
Kinzler Kenneth W.
Papadopoulos Nickolas
Vogelstein Bert
Banner & Witcoff , Ltd.
Fredman Jeffrey
The Johns Hopkins University
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