Species identification based on the point mutations of histo-blood group ABO genes by PCR-RFLP and direct sequencing

Abstract

Using the PCR-restriction fragment length polymorphism (RFLP) and direct sequencing methods, 181 bp DNA fragments at the ABO blood group locus of human and eight different primates were examined. Through PCR amplification and digestion of the product with Hha-1 restriction enzyme, each of the amplified fragments of human, chimpanzee and green monkey was cut into two fragments of 147 and 34 bp, and the corresponding fragment of the other primates was digested into three fragments of 115, 34 and 32 bp. The 181 bp fragments of chimpanzee and green monkey were digested with Mva-1 into three fragments of 82, 58 and 41 bp, and 69, 58 and 54 bp, while the fragments of human and the other primates were separated into two fragments of 123 and 58 bp. Thus, using Hha-1 and Mva-1, human PCR-amplified product of a part of the ABO gene could be discriminated from those of the other primates examined. In addition, by direct sequencing of the 181 bp DNA fragment, two Mva-1 recognition sites and one Hha-1 recognition site were found in the fragments of chimpanzee and green monkey, one Mva-1 site and one Hha-1 site were detected in humans, and one Mva-1 site and two Hha-1 sites in the other primates. These results corresponded well with the data of PCR-RFLP. This method has a good potential for species identification at the DNA level.