Selectable surface marker for sorting transfected B cells

Michael Newton Robertson newton at saul
Sun Feb 26 13:27:41 EST 1995


Why not use human CD4?  A number of expression vectors are available, 
such as pLT4SN which has hCD4 in the retroviral vector pLXSN.  We have 
used this to transduce human and macaque B cell lines, then select for 
transduced cells by using commercially available anti-CD4 coated flasks.
The only problem may be that CD4 itself has some influence on the cell 
cycle, but without a ligand or cross-linking, this should be unlikely.

I have also expressed F-MuLV Env in EL4 cells via a retroviral vector and 
selected high producers by cell sorting using an anit-Env antibody.  This 
is not commercially available, but it can be obtained.  Let me know if 
you are interested.


Michael N. Robertson, MD  Telephone (206) 685-3666 FAX 685-3639
Department of Medicine, Division of Infectious Diseases, SC-42
Retrovirus Lab, Room T-293A, University of Washington Medical Center
1959 NE Pacific St, Seattle, WA  98195  USA

On 22 Feb 1995, Thomas Chiles wrote:

> 
> We are conducting transient transfections in murine Bal-17 B cells to
> examine the effects of antisense mRNAs on cell cycle.  However, only about
> 20 % of the cells take up the antisense plasmid and there is a certain
> amount of read-through from the non-transfected cells.  Although the
> background is not a major problem in our experiments, we would like to
> separate the transfected cells from the nontransfected cells. Is anyone
> aware of an expression vector encoding a surface antigen that is not
> normally expressed on murine B cells which we can co-transfect with the
> antisense construct and then select for surface expession via panning of
> FACS?
> 
> Brian Mcmanus
> Boston College
> at ChilesT at hermes.bc.edu
> 
> 



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