ASSESSMENT OF ENVIRONMENTAL STRESS USING GENE EXPRESSION IN AQUATIC INSECT

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In order to develop the methods assessing environmental stress using gene expression of aquatic midges, the stress-induced or insecticide-induced gene fragments were isolated and gene expression was examined in insecticide-sensitive and field strains of Chironomus yoshimatsui1) Assessment of environmental stress by expression of stress-induced genesThe gene fragment of heat-shock protein, hsc70 was isolated from mRNA of field strain of the midge by RT-PCR reaction using the primer designed from hsp70 base sequences of four species insects and used for Northern blot analysis. In order to assess environmental stresses, RNA isolated from field strains were hybridized with hsc70 cDNA. The expression of this gene was higher in the midges collected in early MayThe inducible gene of hsp70 was also isolated using RNA from heat-treated midges by PT-PCR reaction using the primer designed from base sequence of Chironomus tentans and determined the sequence of full length gene. Using the probe specific for inducible gene, the expression of hsp70 was examined in RNA of heat-treated midges and revealed to reach the peak within 1hour and decline 6 hours later. These results show that the isolated hsp70 gene is inducible2) Assessment of gene expression induced by insecticide-treatmentIn order to assess insecticide-stress, RNAs were isolated from insecticide-treated midges and gene fragments increased by insecticide-treatment were separated in differential display method. Four fragments were isolated. The expression of these gene were higher in insecticide-treated midges and in field strains