Article

A successful strategy for comparative mapping with human ESTs: 65 new regional assignments in the pig.

INRA, Laboratoire de Génétique Cellulaire, BP27, 31326 Castanet-Tolosan Cedex, France.
Mammalian Genome (impact factor: 2.89). 03/1999; 10(2):145-53.
Source: PubMed

ABSTRACT Large-scale sequencing of cDNAs from numerous tissues is currently being performed within the framework of the Human Genome Project. These expressed sequence tags (ESTs) are then mapped on a radiation hybrid panel to produce a high-resolution map of human genes. In this report, we estimate the efficiency of mapping these ESTs in the pig. A total of 344 human ESTs from Généthon were selected for amplification in other species by Zoo-PCR: 186 of these could be reproducibly amplified by use of pig DNA and the corresponding human primer pairs. One-hundred seven of these were tested on a porcine-rodent somatic cell hybrid panel, permitting regional localizations of 65 ESTs with agarose or single-strand conformation polymorphism analysis gels. The corresponding pig PCR products were sequenced: 60 ESTs matched significantly with the expected human sequences. Fifty-one of these localizations in the pig are in agreement with the comparative mapping data between humans and pigs based on heterologous chromosome painting. Seven ESTs that were localized in an unexpected region may indicate new chromosomal correspondences. This work significantly increases the number of genes mapped on the pig genome and demonstrates that this approach can be successfully applied to improve the gene density of mammalian genomic maps in chromosomal regions of interest, such as those in which QTL (Quantative Trait Loci) have been identified.

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Keywords

344 human ESTs
 
corresponding human primer pairs
 
corresponding pig PCR products
 
expected human sequences
 
expressed sequence tags
 
gene density
 
genes mapped
 
heterologous chromosome painting
 
human genes
 
Human Genome Project
 
Large-scale sequencing
 
mammalian genomic maps
 
new chromosomal correspondences
 
pig DNA
 
pig genome
 
pigs
 
porcine-rodent somatic cell hybrid panel
 
Quantative Trait Loci
 
unexpected region
 
Zoo-PCR