The cosmid H3.5, containing genes mapping to the murine H-2 Qa region, was used to transfect L cells by the calcium phosphate co-precipitation method. The resultant transfected cells expressed a Qa-like determinant as detected by an immune serum raised against the transfectant cells and Qa specific monoclonal antibodies. Two-dimensional gel analysis revealed the expression of a class I-like heavy chain with a similar molecular mass to the Qa2 antigens of the positive strain B10 and B10.A but with a different isoelectric point. The cosmid H3.5 spans 40 kb of DNA and contains at least one complete Qa region gene which encodes the Qa-like determinant detected in this study.