A high-resolution cytogenetic-based physical map and a genetic linkage map of human chromosome 16 have been developed based on 79 PCR-typable genetic markers and 2 Southern-based RFLP markers. The PCR-based markers were previously characterized polymorphic (AC)n repeats. Two approaches have led to the characterization of 47 highly informative genetic markers spread along chromosome 16, some of which are closely linked to disease loci. In addition, 22 markers (D16S401-423) previously genetically mapped were also physically mapped. Ten markers characterized by other laboratories were physically mapped and genotyped on the CEPH families. These 32 markers were incorporated into the PCR-based map. Seventy-two markers have heterozygosities > 0.50 and 51 of these markers > 0.70. By multipoint linkage analysis a framework genetic map and a comprehensive genetic map were constructed. The length of the sex-averaged framework genetic map is 152.1 cM. The average distance and the median distance between markers on this map are 3.2 and 2.7 cM, respectively, and the largest gap is 15.9 cM. These maps were anchored to the high-resolution cytogenetic map (on average 1.5 Mb per interval). Together these integrated genetic and physical maps of human chromosome 16 provide the basis for the localization and ultimately the isolation of disease genes that map to this chromosome.

A panel of 54 mouse/human somatic cell hybrids, each possessing various portions of chromosome 16, was constructed; 46 were constructed from naturally occurring rearrangements of this chromosome, which were ascertained in clinical cytogenetics laboratories, and a further 8 from rearrangements spontaneously arising during tissue culture. By mapping 235 DNA markers to this panel of hybrids, and in relation to four fragile sites and the centromere, a cytogenetic-based physical map of chromosome 16 with an average resolution of 1.6 Mb was generated. Included are 66 DNA markers that have been typed in the CEPH pedigrees, and these will allow the construction of a detailed correlation of the cytogenetic-based physical map and the genetic map of this chromosome. Cosmids from chromosome 16 that have been assembled into contigs by use of repetitive sequence fingerprinting have been mapped to the hybrid panel. Approximately 11% of the euchromatin is now both represented in such contigs and located on the cytogenetic-based physical map. This high-resolution cytogenetic-based physical map of chromosome 16 will provide the basis for the cloning of genetically mapped disease genes, genes disrupted in cytogenetic rearrangements that have produced abnormal phenotypes, and cancer breakpoints.

A cosmid contig physical map of human chromosome 16 has been developed by repetitive sequence finger-printing of ∼4000 cosmid clones obtained from a chromosome 16-specific cosmid library. The arrangement of clones in contigs is determined by (1) estimating cosmid length and determining the likelihoods for all possible pairwise clone overlaps, using the fingerprint data, and (2) using an optimization technique to fit contig maps to these estimates. Two important questions concerning this contig map are how much of chromosome 16 is covered and how accurate are the assembled contigs. Both questions can be addressed by hybridization of single-copy sequence probes to gridded arrays of the cosmids. All of the fingerprinted clones have been arrayedon nylon membranes so that any region of interest can be identified by hybridization. The hybridization experiments indicate that ∼84% of the euchromatic arms of chromosome 16 are covered by contigs and singleton cosmids. Both grid hybridization (26 contigs) and pulsed-field gel electrophoresis experiments (11 configs) confirmed the assembled contigs, indicating that false positive overlaps occur infrequently in the present map. Furthermore, regional localization of 93 contigs and singleton cosmids to a somatic cell hybrid mapping panel indicates that there is no bias in the coverage of the euchromatic arms.

Mapping of 33 anonymous DNA probes and 12 genes to the long arm of chromosome 16 was achieved by the use of 14 mouse/human hybrid cell lines and the fragile site FRA16B. Two of the hybrid cell lines contained overlapping interstitial deletions in bands q21 and q22.1. The localization of the 12 genes has been refined. The breakpoints present in the hybrids, in conjunction with the fragile site, can potentially divide the long arm of chromosome 16 into 16 regions. However, this was reduced to 14 regions because in two instances there were no probes or genes that mapped between pairs of breakpoints.

TNFR1 and TNFR2, the genes encoding the two forms of the human tumor necrosis factor receptor, were localized to normal human chromosomes by in situ hybridization and Southern blot analysis of a series of human x mouse hybrid cell lines. TNFR1 maps to 12p13 and TNFR2 maps to 1p36.

Journal Article An STS at the D16S290 locus Get access L.Z Chen, L.Z Chen Department of Cytogenetics and Molecular Genetics, Adelai00de Children's HospitalNorth Adelaide, South Australia 5006, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Y. Shen, Y. Shen Department of Cytogenetics and Molecular Genetics, Adelai00de Children's HospitalNorth Adelaide, South Australia 5006, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar K. Holman, K. Holman Department of Cytogenetics and Molecular Genetics, Adelai00de Children's HospitalNorth Adelaide, South Australia 5006, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar A. Thompson, A. Thompson Department of Cytogenetics and Molecular Genetics, Adelai00de Children's HospitalNorth Adelaide, South Australia 5006, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar S. Lane, S. Lane Department of Cytogenetics and Molecular Genetics, Adelai00de Children's HospitalNorth Adelaide, South Australia 5006, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar R.I. Richards, R.I. Richards Department of Cytogenetics and Molecular Genetics, Adelai00de Children's HospitalNorth Adelaide, South Australia 5006, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar G.R. Sutherland, G.R. Sutherland Department of Cytogenetics and Molecular Genetics, Adelai00de Children's HospitalNorth Adelaide, South Australia 5006, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar D.F. Callen D.F. Callen Department of Cytogenetics and Molecular Genetics, Adelai00de Children's HospitalNorth Adelaide, South Australia 5006, Australia Search for other works by this author on: Oxford Academic PubMed Google Scholar Nucleic Acids Research, Volume 19, Issue 20, 25 October 1991, Page 5793, https://doi.org/10.1093/nar/19.20.5793 Published: 25 October 1991 Article history Received: 13 August 1991 Published: 25 October 1991

A mouse/human hybrid cell panel of human chromosome 16 has been extended to a total of 31 hybrids. These hybrids were derived from constitutional translocations and deletions ascertained during clinical cytogenetic studies. This panel of hybrids, together with four fragile sites, have the potential to divide chromosome 16 into 38 regions. Rapid detailed physical mapping of gene probes or anonymous DNA probes is possible using this hybrid panel. This hybrid cell panel also allows the physical mapping of other chromosomes with three breakpoints on chromosomes 1, 4, 11 and 13 and two on chromosomes 3, 10 and 18.