Expression of the 32-Kd polypeptide of the Rh antigen

Kimita Suyama, Samar Roy, Ruth Lunn, Jack Goldstein

Research output: Contribution to journalArticlepeer-review

15 Scopus citations


To develop procedures for the expression of Rh antigencontaining proteins, we prepared a previously cloned Rh cDNA by isolation from a human bone marrow library using polymerase chain reaction (PCR). The cDNA that encodes either Rh(E) or (e) antigens was inserted into a vector (pGEM7ZRh) and transcription/translation performed in a reticulocyte lysate system. This produced a major (32 Kd) and minor (30 Kd) protein band in the region of sodium dodecyl sulfate (SDS) gels where Rh proteins are known to migrate. These same two proteins were immunoprecipitated by a rabbit anti-Rh antibody preparation that reacts with all Rh proteins, confirming that Rh protein was expressed in this cell-free system. Transient expression of Rh protein in COS-1 cells was also accomplished using a transfection vector (pBC12BIPLRh) containing inserted Rh cDNA. COS-1 cells transfected with this vector also produced a 32-Kd protein band that formed an immune complex with rabbit anti-Rh, while cells transfected with the same vector minus the Rh cDNA insert did not yield any detectable Rh immune-complexed protein. This system should prove useful for studying the transport of Rh proteins within the cell and the expression of Rh antigenicity at the cell surface.

Original languageEnglish (US)
Pages (from-to)1006-1009
Number of pages4
Issue number3
StatePublished - Aug 1 1993
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology


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