Fig. 2

Location and detection of CCR5 sequence polymorphisms

(A) Schema of the CCR5 ORF. Regions of the 1056 nucleotide ORF encoding transmembrane domains I-VII are shown in black, the proposed extracellular domains in white, and the intracellular domains are stippled in the linear scheme shown. The sites of the 32 nucleotide deletion (Δ32) and a Bgl II restriction site are indicated. The double arrow indicates the position of the premature stop codon (TGA) found in CCR5-2. (B) Resolution of CCR5 alleles by restriction fragment length polymorphism analysis (top) and allele-specific PCR ELISA (bottom). For both analyses, the samples studied are indicated above the corresponding column coded as follows: M, 100 base pair interval markers; CCR5-1 and CCR5-2, plasmids containing CCR5-1 (wild type) and CCR5-2 (mutant) ORFs respectively; 1–3, individual human genomic DNA samples. Genotypes are noted below the corresponding column of samples and coded as follows: C1 and C2 are the corresponding plasmid controls; 1/1, CCR5-1 homozygote; 1/2, CCR5-1/CCR5-2 heterozygote; 2/2, CCR5-2 homozygote. (Top) CCR5 amplicons were digested with Bgl II and analyzed by agarose gel electrophoresis. At the right are the lengths (in base pairs) of the indicated fragments. At the left are the lengths in base pairs of the marker standards. (Bottom) The ELISA-based PCR detection of the same samples. The allele specific oligonucleotide probes from CCR5-1 and CCR5-2 (5.1 and 5.2, respectively) are indicated to the left of the corresponding row of samples.