A powerful new technique for isolating genes encoding cell surface antigens using retroviral expression cloning

Andrew C W Zannettino, John R. Rayner, Leonie K. Ashman, Thomas J. Gonda, Paul J. Simmons

Research output: Contribution to journalArticle

41 Citations (Scopus)

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 retrovital 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 β1 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.

Original languageEnglish
Pages (from-to)611-620
Number of pages10
JournalJournal of Immunology
Volume156
Issue number2
Publication statusPublished - 15 Jan 1996
Externally publishedYes

ASJC Scopus subject areas

  • Immunology

Cite this