A POWERFUL NEW TECHNIQUE FOR ISOLATING GENES ENCODING CELL-SURFACE ANTIGENS USING RETROVIRAL EXPRESSION CLONING

cDNA expression cloning using retroviral vectors provides a means of s tably 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 wi de range of cell types, including cells of hemopoietic origin, Here we report a novel method for rapidly isolating genes encoding cell surfa ce 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 Ah-coated magnetic beads, we have successfully isolated six cDNA encoding previously defi ned CSM, including beta(1) integrin and endoglin. Moreover, we have us ed this approach to define the gene and hence the CSM identified by th ree previously unclustered mAb, These results confirm previous studies demonstrating the general utility of retroviral cDNA libraries and fu rther extend their use to the expression cloning of cDNA encoding CSM.