Complementary DNA sequencing: expressed sequence tags and human genome project

  title={Complementary DNA sequencing: expressed sequence tags and human genome project},
  author={Mark Adams and Jm Kelley and J. D. Gocayne and Mark Dubnick and M. M. Polymeropoulos and H. Xiao and CR Merril and And Wu and Björn Olde and RF Moreno and al. et},
  pages={1651 - 1656}
Automated partial DNA sequencing was conducted on more than 600 randomly selected human brain complementary DNA (cDNA) clones to generate expressed sequence tags (ESTs). ESTs have applications in the discovery of new human genes, mapping of the human genome, and identification of coding regions in genomic sequences. Of the sequences generated, 337 represent new genes, including 48 with significant similarity to genes from other organisms, such as a yeast RNA polymerase II subunit; Drosophila… 

Functional genomics in mice by tagged sequence mutagenesis

A gene trap retrovirus shuttle vector has been developed to disrupt genes expressed in murine embryonic stem (ES) cells, and tagged sequence mutagenesis suggests that many of the 10,000–20,000 genes expressing in ES cells can be targeted, providing defined mutations for the analysis of gene functions in vivo.

Sequence identification of 2,375 human brain genes

2,672 new, independent cDNA clones isolated from four human brain cDNA libraries are partially sequenced to generate 2,375 expressed sequence tags to nuclear-encoded genes, representing an approximate doubling of the number of human genes identified by DNA sequencing and may represent as many as 5% of the genes in the human genome.

Long human-mouse sequence alignments reveal novel regulatory elements: a reason to sequence the mouse genome.

The utility of sequencing entire genomes of bacteria and fungi is amply demonstrated, and efforts to determine partial sequences of normalized cDNA libraries have generated rich and very useful databases, such as the TIGR database (TDB) and dbEST.

Methods and Utility of EST and Whole Genome Sequencing

Gene targeted genomic sequencing approaches, like methylation filtration, overcome some of the limitations of EST sequencing and constitute an affordable alternative for plant genomic sequencing.

An integrated approach for identifying and mapping human genes.

The feasibility of individual steps of the method is described using the factor IX (F9) gene as a model system and the mapping of several expressed sequences corresponding to a 330-kb YAC containing DNA from human chromosome 6p21 is presented.

Large-scale concatenation cDNA sequencing.

The authors' data indicate that full-length cDNA insert sequences provide significantly more nucleic acid and protein sequence similarity matches than expressed sequence tags (ESTs) for database searching.

2006 expressed-sequence tags derived from human chromosome 7-enriched cDNA libraries.

These studies provide > 2000 ESTs highly enriched for chromosome 7 gene sequences, 180 new chromosome 7 STSs corresponding to ESTs, and a definitive demonstration of the ability to enrich for chromosome-specific cDNAs by direct selection.

Rapid and efficient cDNA library screening by self-ligation of inverse PCR products (SLIP)

A PCR-based method for directed screening of plasmid cDNA libraries and its utility in a screen of libraries used in the Drosophila EST projects is demonstrated, as well as the relative merits of directed cDNA library screening and RT–PCR approaches.

Progress Toward a Transcript Map of the Human Genome

The combination of data on gene expression and putative gene functions inferred from sequence similarity provides a powerful means of assessing the transcriptional activity of the genome in the cells and tissues of an organism.



Closely related transcripts encoded by the neurogenic gene complex enhancer of split of Drosophila melanogaster.

The deduced protein products of m5, m7 and m8 exhibit extensive sequence homology with each other, and all three encode a sequence similar to one of the conserved domains of representatives of the vertebrate myc gene family which is also present in the deducedprotein sequences of the Drosophila achaete‐scute gene complex.

An 'equalized cDNA library' by the reassociation of short double-stranded cDNAs.

  • M. S. Ko
  • Biology, Chemistry
    Nucleic acids research
  • 1990
A library of mouse fibroblastoid Ltk- cells with nearly equal representations of cDNA clones is tried to make, indicating the usefulness of the current procedure for making equalized cDNA libraries.

A new troponin T and cDNA clones for 13 different muscle proteins, found by shotgun sequencing

Cl clones for 13 of the 19 known muscle-specific proteins were identified, in addition to the clone for the new troponin T isotype, and over the region of nucleotide sequence overlap in the tropon in T clones, the new isotype diverges significantly from its counterpart10.

RNA splice junctions of different classes of eukaryotes: sequence statistics and functional implications in gene expression.

A striking similarity among the rare splice junctions which do not contain AG at the 3' splice site or GT at the 5'splice site indicates the existence of special mechanisms to recognize them, and that these unique signals may be involved in crucial gene-regulation events and in differentiation.

A common language for physical mapping of the human genome.

The polymerase chain reaction (PCR), a method that has only come into widespread use during the past 2 years, seems to offer a path toward a physical map that largely circumvents two problems that were prominent in the NRC Committee's discussions.

Construction of a uniform-abundance (normalized) cDNA library.

Libraries of these fragments are suitable for cDNA subtraction, screening, or selection by hybridization and make it possible to detect and analyze cDNA corresponding to species of mRNA present at a low level in a small fraction of the cells in a complex tissue.

Mapping and sequencing the human genome.

  • V. McKusick
  • Biology
    The New England journal of medicine
  • 1989
IN a 1986 editorial, Renato Dulbecco1 proposed that the best way to speed solution of the fundamental problems of cancer was to sequence the human genome completely — that is, to determine the

mRNA in the mammalian central nervous system.

Tremendous progress has been made in understanding the structures and biological properties of macromolecules, largely due to advances brought about by molecular biologists studying bacterial phage and animal viruses.