Confirming 18S rDNA Polymorphism among Helicosporidium Isolates using Restriction Fragment Length Polymorphism (RFLP) Analysis

The non-photosynthetic pathogenic algae Helicosporidiunl show promising potential as a form of biological control against mosquito larvae. By studying the life cycle and modes of transmission of Helicosporidium, more can be determined about their potential effectiveness as a natural pest control agent. Isolates of Helicosporidium DNA collected from different invertebrate hosts (black fly, weevil, bark beetle, mosquito, and collembola) were predicted to have unique phylotypes. To confirm these differences, Helicosporidium 18S rDNA genes were amplified via Polymerase Chain Reaction (PCR) and purified using the QIAGEN PCR purification kit. The DNA fragments were digested using the restriction enzyme Fokl, and a restriction fragment length polymorphism (RFLP) analysis was conducted using gel electrophoresis. The results show that the Helicosporidium isolates found in different host insects do exhibit 18S rDNA polymorphism. This technique helps to visualize and confirm the newly discovered polymorphism among Helicosporidium