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Type: Journal article
Title: A powerful new technique for isolating genes encoding cell surface antigens using retroviral expression cloning
Author: Zannettino, A.
Rayner, J.
Ashman, L.
Gonda, T.
Simmons, P.
Citation: Journal of Immunology, 1996; 156(2):611-620
Issue Date: 1996
ISSN: 0022-1767
Statement of
Andrew C. W. Zannettino, John R. Rayner, Leonie K. Ashman, Thomas J. Gonda and Paul J. Simmons
Abstract: cDNA expression cloning using retroviral vectors provides a means of stably introducing genes into target cells at efficiencies that surpass those achieved by transfection. Furthermore, retroviral vectors allow for the introduction and expression of complex cDNA libraries in a wide range of cell types, including cells of hemopoietic origin. Here we report a novel method for rapidly isolating genes encoding cell surface molecules (CSM) from a human bone marrow stromal cell cDNA library constructed in the retroviral vector, pRUFneo. With a newly described, highly efficient selection strategy using mAb and Ab-coated magnetic beads, we have successfully isolated six cDNA encoding previously defined CSM, including beta 1 integrin and endoglin. Moreover, we have used this approach to define the gene and hence the CSM identified by three previously unclustered mAb. These results confirm previous studies demonstrating the general utility of retroviral cDNA libraries and further extend their use to the expression cloning of cDNA encoding CSM.
Keywords: Connective Tissue
Bone Marrow Cells
Hematopoietic Stem Cells
Cell Line
Membrane Proteins
Recombinant Fusion Proteins
DNA, Complementary
Antibodies, Monoclonal
Antigens, Surface
Immunomagnetic Separation
Cloning, Molecular
Polymerase Chain Reaction
Gene Expression
Base Sequence
Gene Library
Genetic Vectors
Molecular Sequence Data
Rights: © 1996 by The American Association of Immunologists
Published version:
Appears in Collections:Aurora harvest
Medicine publications

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