GM10027
Somatic cell hybrid
Description:
ADDITIONAL HUMAN/CHINESE HAMSTER SOMATIC CELL HYBRIDS
|
Repository
|
NIGMS Human Genetic Cell Repository
|
| Subcollection |
Hybrids |
|
Cell Type
|
Somatic cell hybrid
|
|
Transformant
|
Untransformed
|
|
Family Member
|
22
|
|
Confirmation
|
Karyotypic analysis and Southern blot hybridization
|
|
ISCN
|
Human-Chinese Hamster somatic cell hybrid retaining human chromosome #22 & Chinese Hamster/human translocations
|
|
Remarks
|
|
| IDENTIFICATION OF SPECIES OF ORIGIN |
Species of Origin Confirmed by Nucleoside Phosphorylase, Glucose-6-Phosphate Dehydrogenase, and Lactate Dehydrogenase Isoenzyme Electrophoresis and by Chromosome Analysis |
| |
| GENE MAPPING & DOSAGE STUDIES - X CHROMOSOME |
DNA from this somatic cell hybrid gave negative results in Southern blot hybridization analyses with probes for Xp22.32 (also Yp11.3), MIC2, and Xq28, DXS15. |
| |
| GENE MAPPING & DOSAGE STUDIES - CHROMOSOME 4 |
DNA from this somatic cell hybrid gave a negative result in Southern blot hybridization analysis with a probe for 4p16.2p15.1, D4S20. |
| |
| GENE MAPPING & DOSAGE STUDIES - CHROMOSOME 6 |
DNA from this somatic cell hybrid gave a negative result in Southern blot hybridization analysis with a probe for 6pterp24, D6S7. |
| |
| GENE MAPPING & DOSAGE STUDIES - CHROMOSOME 8 |
DNA from this somatic cell hybrid gave a negative result in Southern blot hybridization analysis with a probe for 8p22p12, NFL. |
| |
| GENE MAPPING & DOSAGE STUDIES - CHROMOSOME 11 |
DNA from this somatic cell hybrid gave positive results in Southern blot hybridization analyses with probes for 11p15.5, D11S12, and 11q21q22, CLG, while no signal was obtained with probes for 11q23.3, ETS1, and 11p13, CAT. It therefore appears that the nonrandom human/rodent translocation chromosomes found in this somatic cell hybrid involve portions of human chromosome #11 translocated to hamster chromosomes. No intact human chromosome #11 was observed cytogenetically. PCR amplification using probes, PCO3 and PCO4, that bridge the upstream (5') start codon of the first exon of human B-globin (11p15.5) provided no evidence for the presence of human chromosome #11. |
| |
| GENE MAPPING & DOSAGE STUDIES - CHROMOSOME 15 |
Ledbetter et al (Genomics 8:614-622,1990) reported that PCR amplification of interspersed repetitive sequences (IRS) using primers directed at human-specific regions of Alu or L1 indicated that this somatic cell hybrid retained portions of human chromosome 15. Confirmation of this result was obtained by fluorescent in situ hybridization. Positive hybridization was obtained for the proximal long arm of human chromosome #15. DNA from this somatic cell hybrid gave a negative result in Southern blot hybridization analysis with a probe for 15q25qter, IGF1R. |
| |
| GENE MAPPING & DOSAGE STUDIES - CHROMOSOME 19 |
Ledbetter et al (Genomics 8:614-622,1990) reported that PCR amplification of interspersed repetitive sequences (IRS) using primers directed at human-specific regions of Alu or L1 indicated that this somatic cell hybrid retained portions of human chromosome 19. Confirmation of this result was obtained by fluorescent in situ hybridization. Positive hybridization was obtained for the short arm of human chromosome #19. |
| |
| GENE MAPPING & DOSAGE STUDIES - CHROMOSOME 21 |
DNA from this somatic cell hybrid gave a negative result in Southern blot hybridization analysis with a probe for 21q22.1q22.2, D21S17. |
| |
| GENE MAPPING & DOSAGE STUDIES - CHROMOSOME 22 |
DNA from this somatic cell hybrid gave a positive result in Southern blot hybridization analysis with a probe for 22q12.3q13.1, PDGFB. |
| |
| Remarks |
Line EyeF3A6; produced by fusing human lymphs with Chinese hamster ovary line K1, adenine requiring auxotroph Ade-1; grown in purine-free F12 medium with 8% macroserum; retains hum chromo #22 (100% of cells) & hum #11/hamster trans chromo |
| Gimbel W, Klein V, Brass N, Fischer U, Piontek K, Overmyer K, Gottert E, Zang KD, Meese E, Isolation and localization of transcribed sequences on human chromosome 22. Cytogenet Cell Genet71:81-5 1995 |
| PubMed ID: 7606934 |
| |
| Saito M, Helin K, Valentine MB, Griffith BB, Willman CL, Harlow E, Look AT, Amplification of the E2F1 transcription factor gene in the HEL erythroleukemia cell line. Genomics25:130-8 1995 |
| PubMed ID: 7774910 |
| |
| Brass N, Fischer U, Mueller HW, Klein V, Meese E, Strategy for chromosomal assignment of expressed sequences derived from heteronuclear RNA. Biotechniques17:88-90, 92 1994 |
| PubMed ID: 7946323 |
| |
| Fukushima A, Okubo K, Sugino H, Hori N, Matoba R, Niiyama T, Murakawa K, Yoshii J, Yokoyama M, Matsubara K, Chromosomal assignment of HepG2 3'-directed partial cDNA sequences by Southern blot hybridization using monochromosomal hybrid cell panels. Genomics22:127-36 1994 |
| PubMed ID: 7959758 |
| |
| Hendershot LM, Valentine VA, Lee AS, Morris SW, Shapiro DN, Localization of the gene encoding human BiP/GRP78, the endoplasmic reticulum cognate of the HSP70 family, to chromosome 9q34. Genomics20:281-4 1994 |
| PubMed ID: 8020977 |
| |
| Gottert E, Klein V, Piontek K, Overmyer K, Zang KD, Meese E, Generation of a chromosome-22-specific c-DNA library as confirmed by FISH analysis. Hum Genet92:623-6 1993 |
| PubMed ID: 8262524 |
| |
| Huebner K, Druck T, LaForgia S, Lasota J, Croce CM, Lanfrancone L, Donti E, Pengue G, La Mantia G, Pelicci PG, et al, Chromosomal localization of four human zinc finger cDNAs. Hum Genet91:217-22 1993 |
| PubMed ID: 8478004 |
| |
| Trofatter JA, MacCollin MM, Rutter JL, Murrell JR, Duyao MP, Parry DM, Eldridge R, Kley N, Menon AG, Pulaski K, et al, A novel moesin-, ezrin-, radixin-like gene is a candidate for the neurofibromatosis 2 tumor suppressor [published erratum appears in Cell 1993 Nov 19;75(4):826] Cell72:791-800 1993 |
| PubMed ID: 8453669 |
| |
| Washington SS, Bowcock AM, Gerken S, Matsunami N, Lesh D, Osborne-Lawrence SL, Cowell J, Ledbetter DH, White RL, Chakravarti A, A somatic cell hybrid map of human chromosome 13. Genomics18:486-95 1993 |
| PubMed ID: 8307557 |
| |
| Frazer KA, Boehnke M, Budarf ML, Wolff RK, Emanuel BS, Myers RM, Cox DR, A radiation hybrid map of the region on human chromosome 22 containing the neurofibromatosis type 2 locus. Genomics14:574-84 1992 |
| PubMed ID: 1427886 |
| |
| Morris SW, Nelson N, Valentine MB, Shapiro DN, Look AT, Kozlosky CJ, Beckmann MP, Cerretti DP, Assignment of the genes encoding human interleukin-8 receptor types 1 and 2 and an interleukin-8 receptor pseudogene to chromosome 2q35. Genomics14:685-91 1992 |
| PubMed ID: 1427896 |
| |
| Budarf ML, McDermid HE, Sellinger B, Emanuel BS, Isolation and regional localization of 35 unique anonymous DNA markers for human chromosome 22. Genomics10:996-1002 1991 |
| PubMed ID: 1680800 |
| |
| Ledbetter SA, Garcia-Heras J, Ledbetter DH, "PCR-karyotype" of human chromosomes in somatic cell hybrids. Genomics8:614-22 1990 |
| PubMed ID: 2276735 |
| |
| Lichter P, Ledbetter SA, Ledbetter DH, Ward DC, Fluorescence in situ hybridization with Alu and L1 polymerase chain reaction probes for rapid characterization of human chromosomes in hybrid cell lines. Proc Natl Acad Sci U S A87:6634-8 1990 |
| PubMed ID: 2395866 |
| |
| Van Keuren ML, Hart IM, Kao FT, Neve RL, Bruns GA, Kurnit DM, Patterson D, A somatic cell hybrid with a single human chromosome 22 corrects the defect in the CHO mutant (Ade-I) lacking adenylosuccinase activity. Cytogenet Cell Genet44:142-7 1987 |
| PubMed ID: 3568763 |
| Split Ratio |
1:4 |
| Temperature |
37 C |
| Percent CO2 |
5% |
| Medium |
Ham's F12 with 2mM L-glutamine or equivalent |
| Serum |
8% fetal bovine serum Not inactivated |
| Substrate |
None specified |
| Subcultivation Method |
trypsin-EDTA |
| Supplement |
- |
|
|