We have made a regional assignment of the type II collagen gene (COL2A1) on human chromosome 12 by means of an in situ hybridization technique with a biotinylated DNA probe. The precise localization of the signal was mapped to the band 12q13.1-q13.2. This result was in agreement with the previous mapping by isotopic in situ hybridization technique (12q13.1-q13.2), but not with the result of Southern hybridization analysis using somatic cell hybrids (12q14.3).
"In situ hybridization. In situ hybridization and rinsing was carried out according to the protocol described by Takahashi et al. (1989, 1990) with slight modification (Inazawa et al., 1991). The hybridization mixture consisted of 0.5 ~g biotinlabeled cDNA, 50~ formamide, 2 • SSC, 10 ~ bovine serum albumin (BSA), and 20~o dextran sulfate (final concentration). "
[Show abstract][Hide abstract] ABSTRACT: We have used a full length cDNA clone (2.2 kb) for the human cytochrome P-450 nifedipine oxidase (CYP3A4) enzyme as a probe to determine its chromosome localization by fluorescencein situ hybridization. CYP3A4 was mapped on R-banded human prometaphase chromosomes, and the precise localization of CYP3A4 on chromosome 7 was further confirmed by a delineation of G-banded pattern on the same prometaphase chromosomes through a combination of UV-filter. We assigned CYP3A4 to chromosome 7 at q22.1.
Journal of Human Genetics 05/1992; 37(2):133-138. DOI:10.1007/BF01899734 · 2.46 Impact Factor
"The probe concentration was 250 ng/20 pl hybridization solution per slide in the present study. In situ hybridization, rinsing , and detection were performed by the methods of Lawrence et al. (1988) and Takahashi et al. (1989). The hybridization solution consisted of denatured DNA in 50% formamide, 2 x SSC, 2 mg BSA/ml (Boehringer), and 10% dextran sulfate (Sigma). "
[Show abstract][Hide abstract] ABSTRACT: To obtain new RFLP markers on human chromosome 11 for a high-resolution map, we constructed a cosmid library from a Chinese hamster x human somatic hybrid cell line that retains only human chromosome 11 in a Chinese hamster genomic background. A total of 3,500 cosmids were isolated by colony hybridization with labeled human genomic DNA. DNA was prepared from 130 of these cosmid clones and examined for RFLP. In 62 of them, polymorphism was detected with one or more enzymes; four RFLPs were VNTR systems. All polymorphic clones were assigned to one of 22 intervals obtained by mapping on a deletion panel of 15 somatic hybrid cell lines containing parts of chromosome 11; 11 clones were finely mapped by in situ hybridization. Although RFLP markers were scattered on the whole chromosome, they were found predominantly in the regions of R-banding. These DNA markers will contribute to fine mapping of genes causing inherited disorders and tumor-suppressor genes that reside on chromosome 11. Furthermore, as one-third of the cosmid clones revealed a band or bands in Chinese hamster DNA, indicating sequence conservation, this subset of clones may be useful for isolating biologically important genes on chromosome 11.
The American Journal of Human Genetics 03/1991; 48(2):258-68. · 10.93 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The human thymidylate synthase (TS) gene was regionally assigned to chromosome band 18p11.32 by nonisotopic in situ hybridization using biotinylated cDNA (1.1 kb insert) and genomic DNA (6.8 kb insert) probes of the human gene. There have been two provisional assignments for the TS gene to 18pter-q12 and 18q21-qter. The present result confirmed the first of these and further localized the TS gene to the telomeric region of the short arm of chromosome 18. The TS gene appears to be a novel telomeric anchor point for the construction of both physical and genetic linkage maps of human chromosome 18.
Human Genetics 11/1990; 85(6):576-80. DOI:10.1007/BF00193577 · 4.82 Impact Factor
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