Genetic Diversity in Helicosporidium Using β-Tubulin as a Marker

The Helicosporidium is a unique pathogen that can infect the larvae of mosquito. Recently, Helicosporidium have been extracted from different invertebrates (black fly, weevil, bark beetle, mosquito and collembola). Observation of nucleic acid sequences will provide insight into the genetic diversity or similarity between the specimens. Analysis of the amino acid make-up will further details of the phylogenetic relationships between all five specimens. Observation and analysis was done by Polymerase Chain Reaction (PCR), gel electrophoresis, and PCR purification. PCR was used by applying known primers to a sample that will specifically bind to the nucleotides encoding for tubulin. These segments were then loaded into an agarose gel, which separated 3 visible bands. These products were then again amplified and purified in order to extract pure tubulin DNA. The newly amplified DNA was again processed by gel electrophoresis and those bands present were then sequenced. Results indicated that the tubulin gene sequences of Helicosporidium found in weevil (Cyrtobagus salviniae), beetle (Dendroctonus micans), and mosquito (Culex nigripalpus) are genetically similar. Comparison of nucleotide sequences showed some variance, while alignment of their corresponding amino acids showed no variance