Multiple, independent restriction site polymorphisms in human DNA detected with a cDNA probe to argininosuccinate...

We have used a cDNA clone of the human urea cycle enzyme argininosuccinate synthetase (AS) to screen for restriction fragment length polymorphisms (RFLPs) using a large panel of restriction enzymes. The probe, pAS-1, detects from 15 to 27 human DNA fragments by Southern gel analysis. In addition to the structural locus on chromosome 9, AS-like sequences are found on at least 10 human chromosomes, including the X and Y. This large number of dispersed pseudogenes accounts for the multiplicity of hybridizing fragments detected with pAS-1. Of 37 restriction enzymes tested, 18 produced excellent digest patterns; of these 18 enzymes, three revealed high-frequency, independent RFLPs, testing a minimum of 16 individuals with each enzyme. The enzymes producing high-frequency polymorphisms are Hind III (allele frequencies .30 and .70), Hind II (.13 and .87), and Bam HI (.56 and .44). Most of the polymorphic alleles are found in Caucasians, American blacks, and Orientals. The RFLP detected with Hind III maps to chromosome 9 (9q11-q22), although not to the structural locus; the others are autosomal but otherwise unassigned. Two additional six-base enzymes and a four-base enzyme, Msp I, revealed further individual variation, but these variants have not been shown to segregate in families yet. We observed approximately 0.8% high-frequency RFLPs per cut site per enzyme or a minimum of .05% heterozygosity per nucleotide. pAS-1 should prove useful in assigning high-frequency RFLPs to several human chromosomes.