Related Experiment Videos
Identification of human chromosome region 3p14.2-21.3-specific YAC clones using Alu-PCR products from a radiation
T S Siden1, J Kumlien, T Drumheller
1Department of Internal Medicine, Wayne State University School of Medicine, Detroit, Michigan 48201.
Abstract:
Deletion of DNA sequences from at least three different regions on the short arm of human chromosome 3 (3p13-14, 3p21 and 3p25) are frequently observed during the development of many solid tumors, including lung cancers and renal cell carcinomas. In order to physically characterize the 3p21 region, we previously identified a radiation fusion hybrid that contained about 20 megabases of DNA from chromosome region 3p14.2-p21.3. In this study total Alu-PCR products from this hybrid were used as a probe to isolate 86 yeast artificial chromosomes (YAC) clones from a 620-kb average insert YAC library (ICRF). Sixty-nine Alu-PCR markers, generated from the YACs, and seven PCR primers were used to screen for overlaps between individual clones. Seven contigs were identified encompassing 32 YAC clones. Based on previous information about localization of the PCR primers, the three largest contigs could be assigned to smaller subregions between 3p14.2 and 3p21.3. By this work a large proportion of the 3p14.2-21.3 region is covered with large-insert YAC clones.
Insights
Deletions in chromosome 3p regions are common in solid tumors. This study physically maps the 3p14.2-21.3 region using yeast artificial chromosomes (YACs), identifying seven contigs covering a significant portion of this cancer-associated area.
Area of Science:
- Genetics
- Cancer Biology
- Molecular Biology
Background:
- Deletions in specific regions of human chromosome 3 short arm (3p), including 3p13-14, 3p21, and 3p25, are frequently observed in various solid tumors.
- These deletions are particularly prevalent in lung cancers and renal cell carcinomas, suggesting critical tumor suppressor genes within these chromosomal areas.
Purpose of the Study:
- To physically characterize the 3p21 chromosomal region.
- To construct a physical map of the 3p14.2-p21.3 region using large-insert yeast artificial chromosome (YAC) clones.
Main Methods:
- Utilized Alu-PCR products from a radiation fusion hybrid containing ~20 megabases of DNA from the 3p14.2-p21.3 region as a probe.
- Screened a yeast artificial chromosome (YAC) library to isolate clones containing DNA from the target region.
- Employed Alu-PCR markers and PCR primers to identify overlaps and assemble YAC clones into contigs.
Main Results:
- Successfully isolated 86 yeast artificial chromosome (YAC) clones from the target region.
- Generated 69 Alu-PCR markers and utilized seven PCR primers to analyze clone overlaps.
- Identified seven distinct contigs, with 32 YAC clones contributing to these assemblies.
- The three largest contigs were assigned to subregions within 3p14.2-p21.3, covering a substantial portion of this genomic area.
Conclusions:
- This research provides a significant physical coverage of the 3p14.2-21.3 chromosomal region using large-insert YAC clones.
- The generated contigs and markers serve as a foundation for further detailed genetic mapping and identification of genes involved in tumor development.
- This physical map aids in understanding the genetic alterations associated with deletions in chromosome 3p during tumorigenesis.