Generation of a gene model for Aedes aegypti perilipin-1 gene and a probe to investigate number and size of the transcripts in mosquito tissues

Abstract

The mosquito Aedes aegypti is a known vector of disease for humans. Lipid metabolism is of key importance for survival and reproduction. Perilipin-1, PLIN1, is an important protein in the mobilization and storage of lipids in insects. Though the genome of A. aegypti has been sequenced, the gene structures must be confirmed by independent methods. Similarly, the nature of the transcripts from the gene must also be confirmed. From the available genomic and transcriptomic information, a cDNA clone was produced and used to obtain antibodies for western blot assays. Preliminary studies in which protein size was ascertained showed a protein much larger than was predicted by the reported sequences and the clones produced in the lab. Further studies which included cloning and sequencing led to the development of cDNA predicting a protein of the size found in the western blot. Using this information and the newly sequenced Aedes albopictus genome, a structural model of the PLIN1 gene has been constructed and a northern blot probe designed and produced to determine the number and size of the transcripts.