Abstract
Using DNA from L cells which expressed high levels of the CD7 (Leu-9 or HuLy-m2) antigen obtained after two cycles of transfection, a genomic library was constructed in the lambda phage Charon 4A. Recombinant clones containing the gene coding for this antigen were identified by first screening the library with both the HSVtk gene and a probe detecting the human repetitive (Alu) sequences. DNA from 10 tk+ and 12 Alu+ recombinant clones was used to transfect L cells which were analyzed for the cell-surface expression of CD7 either early (48–72 h posttransfection) or later when hypoxanthine aminopterin thymidine-resistant colonies were obtained. Transfection with either Alu+ or tk+ recombinant phages led to transient early expression of CD7, and stable CD7+ transfectants were also established. Thus the CD7 gene has been isolated in a number of clones in association with either the Alu repetitive sequence or with the HSV-tk gene; the insert size in one of the genomic clones was 13.5 kb.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Benton, W. D. and Davis, R. W.: Screening λgt recombinant clones by hybridisation to single plaques in situ. Science 196: 180–182, 1977
Chang, L. J. -A., Gamble, C. L., Izaguirre, A., Minden, M. D., Mak, T. W., and McCulloch, E. A.: Detection of genes coding for human differentiation markers by their transient expression after DNA transfer. Proc. Natl. Acad. Sci. U.S.A. 79: 146–150, 1982
Cosgrove, L. J., Sandrin, M. S., Rajasekariah, P., and McKenzie, I. F. C.: Description of a single genomic clone coding for the β chain of the OKM1, LFA-1 and platelet IIb-IIIa molecules. Proc. Natl. Acad. Sci. U.S.A. 83: 752–756, 1986
Graham, F. L. and Van der Eb, A. J.: A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52: 456–467, 1973]
Gusella, J. F., Keys, C., Varsanyi-Breiner, A., Kao, F. T., Jones, C., Puck, T. T., and Housman, D.: Isolation and localisation of DNA segments from specific human chromosomes. Proc. Natl. Acad. Sci. U.S.A. 77: 2829–2833, 1980
Hayes, B. F., Eisenbarth, G. S., and Rauci, A. S.: Human lymphocyte antigens: Production of a monoclonal antibody that defines functional thymus-derived lymphocyte subsets. Proc. Nod. Acad. Sci. U.S.A. 76: 5829–5833, 1979
Kavathas, P. and Herzenberg, L. A.: Stable transformation of mouse L cells from human membrane T-cell differentiation antigens, HLA and β2 microglobulin: Selection by fluorescence-activated cell sorting. Proc. Natl. Acad. Sci. U.S.A. 80: 524–528, 1983a
Kavathas, P. and Herzenberg, L. A.: Amplification of a gene coding for human T-cell differentiation antigen. Nature 306: 385–387, 1983b
Kavathas, P., Sukhatma, V. P., Herzenberg, L. A., and Parties, J. R.: Isolation of the gene encoding the human T-lymphocyte differentiation antigen Leu-2 (T8) by gene transfer and cDNA subtraction. Proc. Natl. Acad. Sci. U.S.A. 81: 7688–7692, 1984
Kuhn, L. C., Barbosa, J. A., Kamarck, M. E., and Ruddle, F. H.: An approach to the cloning of cell surface protein genes: Selection by cell sorting of mouse L-cells that express HLA or 4172 antigens after transformation with total human DNA. Mol. Biol. Med. 1: 335–352, 1983
Kuhn, L. C., McClelland, A., and Ruddle, F. H.: Gene transfer expression and molecular cloning of the human transferrin receptor gene. Cell 37: 95–103, 1984
Lin, P. F., Zhao, S. Y., and Ruddle, F. H.: Genomic cloning and preliminary characterization of the human thymidine kinase gene. Proc. Natl. Acad. Sci. U.S.A. 80: 6528–6532, 1983
Link, M., Warnke, R., Finlay, J., Amylon, M., Miller, J., Dilley, J., and Levy, R.: A single monoclonal antibody identifies T-cell lineage of childhood lymphoid malignancies. Blood 62: 722–728, 1983
Littman, D. R., Thomas, Y., Maddon, P. J., Chess, L., and Axel, R.: The isolation and sequence of the gene encoding T8: A molecule defining functional classes of T lymphocytes. Cell 40: 237–246, 1985
Maddon, P. J., Littman, D. R., Godfrey, M., Maddon, D. E., Chess, L., and Axel, R.: Isolation and nucleotide sequence of a cDNA encoding the T cell surface protein T4: A new member of the immunoglobulin gene family. Cell 42: 93–104, 1985
Maniatis, T., Hardison, R. C., Lacy, E., Lauer, J., O'Connell, C., Quon, D., Sim, G. K., and Efstratiadis, A.: Thei solation of structural genes from libraries of eucaryotic DNA. Cell 15: 687–701, 1978
Newman, R., Domingo, D., Trotter, J., and Trowbridge, L: Selection and properties of a mouse L-cell transformant expressing human transferrin receptor. Nature 304: 643–645, 1983
Parish, C. R. and McKenzie, I. F. C.: A sensitive ro setting method for detecting subpopulations of lymphocytes which react with alloantisera. J. Immunol. Methods 20: 173–184, 1978
Perucho, M., Hanahan, D., and Wigler, M.: Genetic and physical linkage of exogeneous sequences in transformed cells. Cell 22: 308–317, 1980
Sandrin, M. S., Lovering, K. E., Tachas, G., Collins, P. R., and McKenzie, I. F. C.: Gene transfection of the HuLy-m2 (Leu-9) antigen into mouse L cells. Immunogenetics 25: 279–283, 1987
Scangos, G. and Ruddle, F. H.: Mechanisms and applications of DNA-mediated gene transfer in mammalian cells - A review. Gene 14: 1–10, 1981
Shih, C. and Weinberg, R. A.: Isolation of a transforming sequence from a human bladder carcinoma cell line. Cell 29: 161–169, 1982
Southern, E.: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98: 503–517, 1975
Thurlow, P. J., Lovering, K. E., and McKenzie, I. F. C.: A monoclonal antibody detecting a new human T cell antigen, HuLy-m2. Transplantation 38: 143–147, 1984
Wigler, M., Pellicer, A., Silverstein, S., and Axel, R.: Biochemical transfer of a single copy eucaryotic gene using total cellular DNA as donor. Cell 14: 725–731, 1978
Wigler, M., Sweet, R., Sim, G. K., Wold, B., Pellicer, A., Lacy, E., Maniatis, T., Silverstein, S., and Axel, R.: Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell 16: 777–785, 1979
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Erin Lovering, K., Deacon, N.J., Mickelson, C.A. et al. Isolation of the CD7 gene from the DNA of transfected L cells. Immunogenetics 25, 391–396 (1987). https://doi.org/10.1007/BF00396105
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00396105