Dataset Information

Radiation hybrid mapping of the two highly homologous human-variant pMCHL genes by PCR-SSCP.

ABSTRACT: When gene loci are very similar in sequence, as in gene families or multiple pseudogenes, it is difficult to determine the specific location of the individual genes. We show here that applying PCR-SSCP to a radiation hybrid panel allowed mapping and specific sequencing of two genes with only a few sequence differences. Human-variant forms of the promelanin-concentrating hormone (pMCH) gene are found in two locations in the genome, previously localized by FISH to 5p14 and 5q12-q13. Without prior knowledge of sequence variation between the loci, we observed a difference in migration pattern in PCR-SSCP, indicating the presence of at least one point of sequence divergence. PCR-SSCP of 93 samples from a human-hamster radiation hybrid panel revealed the location of the genes to be between markers WI-4804 and AFM225YC5 on chromosome 5p, and between markers WI-3133 and WI-4225 on chromosome 5q. Sequencing of the two 680-bp PCR products from the hybrid panel demonstrated 3 bases of sequence difference between the 5p and 5q locations.

SUBMITTER: Miller CL

PROVIDER: S-EPMC310753 | biostudies-literature | 1998 Jul

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Radiation hybrid mapping of the two highly homologous human-variant pMCHL genes by PCR-SSCP.

Miller C L CL Thompson R C RC Burmeister M M

Genome research 19980701 7

When gene loci are very similar in sequence, as in gene families or multiple pseudogenes, it is difficult to determine the specific location of the individual genes. We show here that applying PCR-SSCP to a radiation hybrid panel allowed mapping and specific sequencing of two genes with only a few sequence differences. Human-variant forms of the promelanin-concentrating hormone (pMCH) gene are found in two locations in the genome, previously localized by FISH to 5p14 and 5q12-q13. Without prior ...[more]

PMID: 9685321

Similar Datasets

Project description:BackgroundBovine chromosome (BTA) 15 contains a quantitative trait loci (QTL) for meat tenderness, as well as several breaks in synteny with human chromosome (HSA) 11. Both linkage and radiation hybrid (RH) maps of BTA 15 are available, but the linkage map lacks gene-specific markers needed to identify genes underlying the QTL, and the gene-rich RH map lacks associations with marker genotypes needed to define the QTL. Integrating the maps will provide information to further explore the QTL as well as refine the comparative map between BTA 15 and HSA 11. A recently developed approach to integrating linkage and RH maps uses both linkage and RH data to resolve a consensus marker order, rather than aligning independently constructed maps. Automated map construction procedures employing this maximum-likelihood approach were developed to integrate BTA RH and linkage data, and establish comparative positions of BTA 15 markers with HSA 11 homologs.ResultsThe integrated BTA 15 map represents 145 markers; 42 shared by both data sets, 36 unique to the linkage data and 67 unique to RH data. Sequence alignment yielded comparative positions for 77 bovine markers with homologs on HSA 11. The map covers approximately 32% of HSA 11 sequence in five segments of conserved synteny, another 15% of HSA 11 is shared with BTA 29. Bovine and human order are consistent in portions of the syntenic segments, but some rearrangement is apparent. Comparative positions of gene markers near the meat tenderness QTL indicate the region includes separate segments of HSA 11. The two microsatellite markers flanking the QTL peak are between defined syntenic segments.ConclusionsCombining data to construct an integrated map not only consolidates information from different sources onto a single map, but information contributed from each data set increases the accuracy of the map. Comparison of bovine maps with well annotated human sequence can provide useful information about genes near mapped bovine markers, but bovine gene order may be different than human. Procedures to connect genetic and physical mapping data, build integrated maps for livestock species, and connect those maps to more fully annotated sequence can be automated, facilitating the maintenance of up-to-date maps, and providing a valuable tool to further explore genetic variation in livestock.

Dataset Information

Radiation hybrid mapping of the two highly homologous human-variant pMCHL genes by PCR-SSCP.

Publications

Radiation hybrid mapping of the two highly homologous human-variant pMCHL genes by PCR-SSCP.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets