A notI RFLP in the human alpha 2-C4 adrenergic receptor locus (ADRA2RL2) detected by PFGE.

Description: The thiolester site of the human alpha-2-macroglobulin (A2M) is essential for the function of A2M as a proteinase inhibitor (1, 2). This site, which is encoded for by exon 24 of the A2M gene, was amplified by polymerase chain reactio (PCRa usinr tho primer 5'-CTGAAGAATTATCCCTGAAAC-3' and 5'-TGGTTCCTGCCATATCGCTC-3'. PCRs were performed in 100 /xl volumes containing 0.5 ng genomic DNA, 25 pmoles of each primer, 200 /iM of each dNTP in 50 mM KC1, 10 mM Tris-HCl, 3.0 mM MgCl2, 0.01% (w/v) gelatin and 2.5 U Taq polymerase. 30 cycles of amplification were conducted, each consisting of 1 min at 94°C, 1 min at 58°C, 30 sec at 72°C. Sequencing of the 0.8 kb amplification product with the primer 5'-ATTAGGCTCTGCCATGCAA-3' according to the method of Wong et al. (3) showed a sequence polymorphism of the gene.