X chromosome usage by hemopoietic and nonhemopoietic cells in a female with CVID and acquired thalassemia analyzed by detection of glucose-6-phosphate dehydration (G6PD) gene polymorphism
(A) The ligase detection analysis (LDR) of a G6PD polymorphism (C/T exonic nucleotide #1311) (8) employed a cDNA prepared by reverse PCR reaction from total cellular RNA (10). The negative control (Neg. Co.) represents a sham experiment without added DNA template. Hair root follicles (H), mouth epithelial cells (Me), skin cultured fibroblasts (S), reticulocytes (R), platelets (P), granulocytes (G), mononuclear cells (M), and T-lymphocytes (T), of a female with CVID (Patient 1) were examined using a cDNA template prepared reaction from total cellular RNA. (B) The LDR analysis employed a positive T/T control (Pos. Co.) reaction with added DNA template from an individual homozygous for the T polymorphism), and as a negative control (Neg. Co.) a sham experiment without added DNA template. B and T lymphocytes were obtained by immunofluorescence sorting of peripheral blood mononuclear cells as described in Methods. Detection of an allelic transcript of an active X chromosome was done as described (8,10).