TY - JOUR
T1 - A powerful new technique for isolating genes encoding cell surface antigens using retroviral expression cloning
AU - Zannettino, Andrew C W
AU - Rayner, John R.
AU - Ashman, Leonie K.
AU - Gonda, Thomas J.
AU - Simmons, Paul J.
PY - 1996/1/15
Y1 - 1996/1/15
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0030060389&partnerID=8YFLogxK
M3 - Article
C2 - 8543812
AN - SCOPUS:0030060389
VL - 156
SP - 611
EP - 620
JO - Journal of Immunology
JF - Journal of Immunology
SN - 0022-1767
IS - 2
ER -