![]() ![]() To analyze the function of an enolase, the gene sequence of the organism of interest is required. These reports suggest that insect enolases can have many different species-specific roles. The expression of an enolase protein was up-regulated in the midgut of Aedes aegypti infected with chikungunya or dengue viruses. ![]() For example, the enolase of the parasitic wasp, Aphidius ervi is expressed on the egg surface and contributes to the digestion of host proteins by promoting plasmin generation as a plasminogen receptor. Insect enolases differ from mammalian enolases in that they have relatively low conservation among insects and show species-specific functions. In recent years, many insect enolases have been discovered. The ENO1 and ENO 3 sequences are well conserved in vertebrates, whereas the insect Enolase 1-like sequence is well conserved across arthropods. A sperm-specific enolase was also identified in Mus musculus. Isoenzyme alpha is present in most tissues, whereas the beta form is localized to the muscle and the gamma form is found only in nervous tissue. In a human study, three enolase isoenzymes were identified as homodimers composed of two alpha (also known as ENO1 Online Mendelian Inheritance in Man (OMIM), 172430), two gamma (ENO2 OMIM, 131360), or two beta (ENO3 OMIM, 131370) subunits. Glycolysis is responsible for the majority of energy production in all organisms. Thus, we need to develop an analysis procedure for how to select functional genes from public databases.Įnolase is a key glycolytic enzyme (2-phospho- d-glycerate hydrolase EC 4.2.1.11). However, many incorrectly annotated genes are included in these databases, which can prevent the correct interpretation of gene annotations in non-model organisms. For our present study, we easily obtained gene sequences and analyses from large datasets, including RNA-seq results, from public databases. Researchers can now access public databases containing the genomic information of many insects for comparative analyses. This project provides genomic information for minor insect species, such as those not used experimentally ( ). The i5k project, which began in 2011, aims to sequence the genomes of 5000 arthropod species. Studies of insects have contributed a wealth of scientific discoveries. Insects can adapt to any number of environmental conditions because of their small size. There are more than one million species of insects in the world. Our strategy could be helpful for the detection of minor genes and functional genes in non-model organisms from public databases. In addition, we determined that BmEno2 and BmEnoC had species-specific functions. We identified and characterized three functional enolases from public databases with a combination of dry and wet bench processes in the silkworm B. Quantitative RT-PCR showed that the mRNA expression of BmEno2 and BmEnoC correlated with testis development thus, BmEno2 and BmEnoC may be related to lepidopteran-specific spermiogenesis. BmEnoC was weakly expressed in the testis. In contrast, BmEno2 was mainly expressed in the testis with some expression in the ovary and suboesophageal ganglion. BmEno1 was expressed in all of the tissues used in our study. Phylogenetic analysis showed that BmEno2 and BmEnoC were distinct from BmEno1 and other enolases, and were distributed only in lepidopteran clusters. However, BmEno2 and BmEnoC showed a change in this key amino acid. BmEno1 contained a conserved key amino acid residue for metal binding and substrate binding in other species. First, we detected five annotated enolases from public databases using a Hidden Markov Model (HMM) search, and then through cDNA cloning, Northern blotting, and RNA-seq analysis, we revealed three functional enolases in B. We used a combination of dry and wet bench processes to identify functional enolases in the silkworm Bombyx mori (BmEno). ResultsĮnolase is an important glycolytic enzyme in all organisms. To address this problem, we used a combination of dry and wet bench processes to select functional genes from public databases. However, many incorrectly annotated genes are included in these databases, which can prevent the correct interpretation of subsequent functional analyses. Thus, researchers can easily obtain online genomic information on invertebrates and insects. Various insect species have been added to genomic databases over the years. ![]()
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