The Human Genome Project, an endeavor to map and sequence the entire human genome, has been in existence for almost seven years. One result of this effort has been the development of increasingly detailed genetic and physical maps spanning large regions of virtually every chromosome. Paralleling this has been the increasingly high resolution mapping of many &wnetic diseases. Together, these developments have facilitated the isolation of specific disease genes and are now motivating the construction of comprehensive transcriptional maps. This latter endeavor represents a new facet of the genome…mehr
The Human Genome Project, an endeavor to map and sequence the entire human genome, has been in existence for almost seven years. One result of this effort has been the development of increasingly detailed genetic and physical maps spanning large regions of virtually every chromosome. Paralleling this has been the increasingly high resolution mapping of many &wnetic diseases. Together, these developments have facilitated the isolation of specific disease genes and are now motivating the construction of comprehensive transcriptional maps. This latter endeavor represents a new facet of the genome project, and as such requires the recognition and solution of a new set of problems, with the attendant development and application of a new set of techniques. The First International Workshop on the Identification of Transcribed Sequences in the Human Genome was held in 1991 and was attended by 23 investigators. Discussions at this meeting were largely devoted to defining the magnitude of the problem and describing the available techniques. A small number of laboratories reported the development of new techniques (at that time, for example, exon trapping, cDNA hybrid selection, direct cDNA screening, use of splice junction conserved sequences,etc.), but data were too limited to permit comparisons of their relative efficiencies.
Introduction: Seven Blind Men and An Elephant.- From Genomic DNA to Transcribed Sequences.- Classical Approaches.- Identification of Genes and Construction of a Transcriptional Map in Xq28.- Use of cDNA Selection and Evolutionarily Conserved Sequences to Isolate Transcribed Sequences from Region Xp11.21.- Identification of cDNAs by Direct Hybridization Using Cosmid Probes.- Hybridization Based Approaches.- Hybridization Selection:.- Locus Specific Identification of Transcribed Sequences Using YACs and Whole Yeast Genomic DNA.- Towards a Transcriptional Map of Human Chromosome 21.- Isolation of Expressed Sequences from the Chromosome 17q21 BRCA1 Region by Magnetic Bead Capture.- Towards a Transcriptional Map of the q21-q22 Region of Chromosome 7.- Direct cDNA Selection Using Human and Mouse cDNAs:Application to Xq13.3 Chromosomal Region.- Variations on Hybridization Selection:.- A Sandwich-Hybridization Method for Specific and Efficient Selection of cDNA Clones from Genomic Regions.- Novel Strategy for Isolating Unknown Coding Sequences from Genomic DNA by Generating Genomic-cDNA Chimeras.- Identifying and Directly Purifying Transcribed Elements by Coincident Sequence Cloning.- Finding Candidate Genes by Preparative in situ Hybridization.- Direct cDNA Screening:.- Direct cDNA Screening of Genomic Reference Libraries - A Rapid Method for the Identification of Transcribed Sequences in Large Genomic Regions.- Identification of Expressed Sequences on Human Chromosome 9q32-34.- Exon Trapping.- An Exon Trapping System Providing Size Selection of Spliced Clones and Facilitating Direct Cloning.- Isolation of Gene Sequences from the BRCA1 Region of Chromosome 17q21 by Exon Amplification.- Isolation of Coding Sequence from Cosmids and YACs by Exon Amplification.- IntegratedTranscriptional Maps of Large DNA Regions: Towards a Transcriptional Map of Human Chromosome 21.- Computer Based Approaches.- Shallow Shotgun Sequencing as a Strategy for Finding Coding Exons.- From Transcribed Sequences to Genomic Localization.- Generating and Sequencing cDNAs.- Requirements in Screening cDNA Libraries for New Genes and Solutions Offered by SBH Technology.- Establishing Catalogues of Expressed Sequences by Oligonucleotide Fingerprinting of cDNA Libraries.- PCR-Based Technologies to Study Differential Gene Expression in Rat Brain.- Mapping cDNAs.- 259 Human Brain Expressed Sequence Tags (ESTs): Chromosome Localization, Subregional Assignment, and Sequence Analysis.- Mapping cDNAs by Hybridization to Gridded Arrays of DNA from YAC Clones.- Discussion.
Introduction: Seven Blind Men and An Elephant.- From Genomic DNA to Transcribed Sequences.- Classical Approaches.- Identification of Genes and Construction of a Transcriptional Map in Xq28.- Use of cDNA Selection and Evolutionarily Conserved Sequences to Isolate Transcribed Sequences from Region Xp11.21.- Identification of cDNAs by Direct Hybridization Using Cosmid Probes.- Hybridization Based Approaches.- Hybridization Selection:.- Locus Specific Identification of Transcribed Sequences Using YACs and Whole Yeast Genomic DNA.- Towards a Transcriptional Map of Human Chromosome 21.- Isolation of Expressed Sequences from the Chromosome 17q21 BRCA1 Region by Magnetic Bead Capture.- Towards a Transcriptional Map of the q21-q22 Region of Chromosome 7.- Direct cDNA Selection Using Human and Mouse cDNAs:Application to Xq13.3 Chromosomal Region.- Variations on Hybridization Selection:.- A Sandwich-Hybridization Method for Specific and Efficient Selection of cDNA Clones from Genomic Regions.- Novel Strategy for Isolating Unknown Coding Sequences from Genomic DNA by Generating Genomic-cDNA Chimeras.- Identifying and Directly Purifying Transcribed Elements by Coincident Sequence Cloning.- Finding Candidate Genes by Preparative in situ Hybridization.- Direct cDNA Screening:.- Direct cDNA Screening of Genomic Reference Libraries - A Rapid Method for the Identification of Transcribed Sequences in Large Genomic Regions.- Identification of Expressed Sequences on Human Chromosome 9q32-34.- Exon Trapping.- An Exon Trapping System Providing Size Selection of Spliced Clones and Facilitating Direct Cloning.- Isolation of Gene Sequences from the BRCA1 Region of Chromosome 17q21 by Exon Amplification.- Isolation of Coding Sequence from Cosmids and YACs by Exon Amplification.- IntegratedTranscriptional Maps of Large DNA Regions: Towards a Transcriptional Map of Human Chromosome 21.- Computer Based Approaches.- Shallow Shotgun Sequencing as a Strategy for Finding Coding Exons.- From Transcribed Sequences to Genomic Localization.- Generating and Sequencing cDNAs.- Requirements in Screening cDNA Libraries for New Genes and Solutions Offered by SBH Technology.- Establishing Catalogues of Expressed Sequences by Oligonucleotide Fingerprinting of cDNA Libraries.- PCR-Based Technologies to Study Differential Gene Expression in Rat Brain.- Mapping cDNAs.- 259 Human Brain Expressed Sequence Tags (ESTs): Chromosome Localization, Subregional Assignment, and Sequence Analysis.- Mapping cDNAs by Hybridization to Gridded Arrays of DNA from YAC Clones.- Discussion.
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