Chromosomal localization and expressed sequence tag generation of clones from a normalized human adult thymus cDNA library.

Abstract

Expressed sequence tags [ESTs) from 298 clones have been generated from a randomly primed, normalized human adult thymus cDNA library. We describe the chromosomal localization of 136 of these ESTs by PCR-based mapping to a human monochromosomal somatic cell hybrid panel. Data base similarities to known genes are also described. A subset {n =18) of these randomly primed ESTs extended the sequence of ESTs from other tissues currently in dbEST. Of the nonrepetitive human adult thymus ESTs generated in this study, 237 (79.5%) have no similarity to current data base entries. This would suggest that our collection contains -100 new coding regions from thymus tissue, a large proportion of which likely will represent the middle regions of genes. The mapped ESTs should prove useful as new gene-based markers for mapping and candidate gene hunting, particularly when anchored to a well-developed physical map of the human genome.

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

“…The single-pass partial sequencing of cDNAs to generate ESTs has proven to be a powerful and successful way to assemble a profile of genes expressed in a particular organism, tissue, or cell type (Adams et al 1991(Adams et al , 1992(Adams et al , 1993a(Adams et al ,b, 1995Khan et al 1992;McCombie et al 1992;Okubo et al 1992;Waterston et al 1992;Hofte et al 1993;Takeda et al 1993;Liew et al 1994;Sudo et al 1994;Frigerio et al 1995;Houlgatte et al 1995;Lamerdin et al 1995;Hillier et al 1996). We have extended this paradigm by coupling direct cDNA selection with EST production to establish a profile of expressed genes from a single human chromosome.…”

“…There were 43 (17%) that did not amplify an appropriate product from h u m a n genomic DNA. This failure rate is typical of efforts to develop STSs from cDNA sequences, with the associated hazards of u n k n o w n intron positions (Hudson et al 1995;Lamerdin et al 1995). There were 12 (5%) that appeared to amplify repetitive sequences, with the appropriate product being generated from some or all h u m a n chromosome-containing cell lines.…”

Genetic Information, Genomic Technologies, and the Future of Drug Discovery

“…13,14 Additional uses of these technologies include the ability to compare normal and disease paradigms of human endothelium by forward and backward cDNA library subtraction technologies and differential display techniques to concentrate on transcripts associated with the disease hypothesis. 15,16 These ap-proaches will provide a high resolution view of the endothelial cell system with thousands of components catalogued as mRNAs of the corresponding proteins.…”