The MDTC-RP30 lymphoblastoid cell line established from Marek's disease (MD) tumors in turkeys consisted of a heterogeneous population of cells 10 to 25 micron in diameter. Large-cell fractions obtained from a bovine fetal serum gradient had a higher titer of cell-associated MD virus (MDV) than the small-cell fractions. Seven single-cell clones were established from MDTC-RP30 cell line: two consisted of large cells, and the other clones consisted of small cells. Infectious MDV was rescued from large-cell clones in chicken embryo fibroblast cultures but not from small-cell clones. All clones contained MDV DNA sequences when hybridized against cloned MDV DNA. All clones were positive for a Marek's-disease-tumor-associated surface antigen and surface immunoglobulins. All but two small-cell clones caused MD in susceptible chickens. The two large-cell-type clones were uniformly tetraploid, whereas one small-cell clone was diploid and the four others were a mixture of diploid and tetraploid, with an occasional triploid cell. Evidence of translocation involving the male (Z) chromosome and the chromosome #3 was seen in one clone. These results suggest that MDV transforms different subpopulations of lymphocytes.