簡便な植物の遺伝子多型解析実験の検討

抄録

  Advances in molecular biology are being made in the fields of science, medicine, and pharmacy. The majority of students graduating from pharmaceutical university departments will be required to have knowledge of molecular biology techniques. Although it is not difficult to perform DNA experiments in a laboratory, they involve a series of operations. Therefore it is difficult to include such experiments in the curriculum for students at our university because of safety concerns and the experimental equipment and time required. This paper introduces a convenient experiment on plant DNA polymorphism using a polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) method that we developed. This experiment enables each student to handle DNA safely. Total DNA is extracted from a piece of leaf derived from Aconitum carmichaeli or Aquilegia flabellate, the nuclear 5.8S rRNA region is amplified by PCR, and the PCR product is digested by restriction enzymes. Different polymorphisms in the plants can be visualized on electrophoregrams. The total time required for the experiments is 2 days (about 4 h per day). In the field of pharmacognosy, authentication of herbal medicine using genetic analysis is increasingly important. Plant DNA polymorphisms required the same knowledge and techniques as genetic analysis and are part of the course content in medicinal plant science. The PCR-RFLP experiment proposed here is a suitable method to acquire molecular biological knowledge and techniques.