Abstract
Mouse teratocarcinoma cells express neither H-2 heavy chains nor beta 2-microglobulin (beta 2-m). We have constructed two genomic libraries, one from PCC4-aza-RI embryonal carcinoma cells and the other from their adult syngenic counterpart 129/Sv liver cells (H-2bc). The libraries were screened with a full length mouse beta 2-m cDNA probe which we isolated and sequenced. Two cosmid clones carrying the entire beta 2-m gene were isolated, one from each library. There was no detectable difference in structure between the two genes. Furthermore, both were shown to be active and to restore beta 2-m synthesis upon transfer into mutant cells deficient in beta 2-m. Irreversible DNA alterations in or around the beta 2-m gene are thus unlikely to account for the lack of beta 2-m gene expression in embryonal teratocarcinoma cells.
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Selected References
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- Berg P. Dissections and reconstructions of genes and chromosomes. Science. 1981 Jul 17;213(4505):296–303. doi: 10.1126/science.6264595. [DOI] [PubMed] [Google Scholar]
- Buc-Caron M. H., Condamine H., Jacob F. The presence of F9 antigen on the surface of mouse embryonic cells until day 8 of embryogenesis. J Embryol Exp Morphol. 1978 Oct;47:149–160. [PubMed] [Google Scholar]
- Croce C. M., Linnenbach A., Huebner K., Parnes J. R., Margulies D. H., Appella E., Seidman J. G. Control of expression of histocompatibility antigens (H-2) and beta 2-microglobulin in F9 teratocarcinoma stem cells. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5754–5758. doi: 10.1073/pnas.78.9.5754. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dubois P., Fellous M., Gachelin G., Jacob F., Kemler R., Pressman D., Tanigaki N. Absence of a serologically detectable association of murine beta2-microglobulin with the embryonic F9 antigen. Transplantation. 1976 Nov;22(5):467–473. doi: 10.1097/00007890-197611000-00009. [DOI] [PubMed] [Google Scholar]
- Gachelin G. The cell surface antigens of mouse embryonal carcinoma cells. Biochim Biophys Acta. 1978 Sep 18;516(1):27–60. doi: 10.1016/0304-419x(78)90003-3. [DOI] [PubMed] [Google Scholar]
- Gross-Bellard M., Oudet P., Chambon P. Isolation of high-molecular-weight DNA from mammalian cells. Eur J Biochem. 1973 Jul 2;36(1):32–38. doi: 10.1111/j.1432-1033.1973.tb02881.x. [DOI] [PubMed] [Google Scholar]
- Hanahan D., Meselson M. Plasmid screening at high colony density. Gene. 1980 Jun;10(1):63–67. doi: 10.1016/0378-1119(80)90144-4. [DOI] [PubMed] [Google Scholar]
- Hohn B., Collins J. A small cosmid for efficient cloning of large DNA fragments. Gene. 1980 Nov;11(3-4):291–298. doi: 10.1016/0378-1119(80)90069-4. [DOI] [PubMed] [Google Scholar]
- Hohn B. In vitro packaging of lambda and cosmid DNA. Methods Enzymol. 1979;68:299–309. doi: 10.1016/0076-6879(79)68021-7. [DOI] [PubMed] [Google Scholar]
- Jakob H., Boon T., Gaillard J., Nicolas J., Jacob F. Tératocarcinome de la spuris: isolement, culture et propriétés de cellules a potentialités multiples. Ann Microbiol (Paris) 1973 Oct;124(3):269–282. [PubMed] [Google Scholar]
- Lalanne J. L., Delarbre C., Gachelin G., Kourilsky P. A cDNA clone containing the entire coding sequence of a mouse H-2Kd histocompatibility antigen. Nucleic Acids Res. 1983 Mar 11;11(5):1567–1577. doi: 10.1093/nar/11.5.1567. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lingappa V. R., Cunningham B. A., Jazwinski S. M., Hopp T. P., Blobel G., Edelman G. M. Cell-free synthesis and segregation of beta 2-microglobulin. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3651–3655. doi: 10.1073/pnas.76.8.3651. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
- Morello D., Daniel F., Baldacci P., Cayre Y., Gachelin G., Kourilsky P. Absence of significant H-2 and beta 2-microglobulin mRNA expression by mouse embryonal carcinoma cells. Nature. 1982 Mar 18;296(5854):260–262. doi: 10.1038/296260a0. [DOI] [PubMed] [Google Scholar]
- Morello D., Gachelin G., Dubois P., Tanigaki N., Pressman D., Jacob F. Absence of reaction of a xenogenic anti-H-2 serum with mouse embryonal carcinoma cells. Transplantation. 1978 Aug;26(2):119–125. doi: 10.1097/00007890-197808000-00012. [DOI] [PubMed] [Google Scholar]
- Murray N. E., Brammar W. J., Murray K. Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet. 1977 Jan 7;150(1):53–61. doi: 10.1007/BF02425325. [DOI] [PubMed] [Google Scholar]
- Natori T., Katagiri M., Tanigaki N., Pressman D. The 11,000-dalton component of mouse H-2. Isolation and identification. Transplantation. 1974 Dec;18(6):550–555. doi: 10.1097/00007890-197412000-00015. [DOI] [PubMed] [Google Scholar]
- POPE J. H., ROWE W. P. DETECTION OF SPECIFIC ANTIGEN IN SV40-TRANSFORMED CELLS BY IMMUNOFLUORESCENCE. J Exp Med. 1964 Aug 1;120:121–128. doi: 10.1084/jem.120.2.121. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Parnes J. R., Seidman J. G. Structure of wild-type and mutant mouse beta 2-microglobulin genes. Cell. 1982 Jun;29(2):661–669. doi: 10.1016/0092-8674(82)90182-9. [DOI] [PubMed] [Google Scholar]
- Rassoulzadegan M., Binetruy B., Cuzin F. High frequency of gene transfer after fusion between bacteria and eukaryotic cells. Nature. 1982 Jan 21;295(5846):257–259. doi: 10.1038/295257a0. [DOI] [PubMed] [Google Scholar]
- Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
- Sawicki J. A., Magnuson T., Epstein C. J. Evidence for expression of the paternal genome in the two-cell mouse embryo. Nature. 1981 Dec 3;294(5840):450–451. doi: 10.1038/294450a0. [DOI] [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Suggs S. V., Wallace R. B., Hirose T., Kawashima E. H., Itakura K. Use of synthetic oligonucleotides as hybridization probes: isolation of cloned cDNA sequences for human beta 2-microglobulin. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6613–6617. doi: 10.1073/pnas.78.11.6613. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wieslander L. A simple method to recover intact high molecular weight RNA and DNA after electrophoretic separation in low gelling temperature agarose gels. Anal Biochem. 1979 Oct 1;98(2):305–309. doi: 10.1016/0003-2697(79)90145-3. [DOI] [PubMed] [Google Scholar]