Molecular Identification Of Ostrinia Furnacalis (guenée) Glutaredoxin Gene

Glutaredoxin, a coenzyme of the ribose nucleotide reductase, is ubiquitous in the organism with dehydroascorbate reductase activity. It plays important roles in regulating the redox reaction of the body, cell growth, and inhibiting apoptosis. In this exp- eriment, the characterizations of glutaredoxin gene from Ostrinia furnacalis were studied and these studies further provide foundation to understand the insect response to adversity, The main results are as follows:（1） Ostrinia furnacalis glutaredoxin （OfurGrx） cDNA was isolated by using reverse transcription polymerase chain reaction （RT-PCR） and rapid amplification of cDNA 3’ends （RACE）. Nucleotide sequence analysis revealed that the cDNA fragment contains a 228 bp putative coding region and 343 bp 3’untranslated region （UTR）. Sequencing analysis showed further that predicted amino acid sequence contains a conserved domains: GRX_GRXh_12like; OfurGrx share 56% identity with Bombyx mori Grx and 9% to 56% with other insects. Altogether, these results suggest that the gene obtained belongs to Grx family genes（GenBank accession number GU393246）.（2） The temporal and tissular expression of OfurGrx was analyzed by using RT-PCR. The results showed that the OfurGrx is expressed in all developmental stages and tissues.（3） The temporal and tissular expression of OfurGrx transcript was studied by using Real-time PCR. The results showed that OfurGrx reach expression peak in the newly -hatched larvae and 4th-instar larvae. Its transcript increased in feeding stages with the growth and decrease in the 4th and 5th instar ecdysis stage, which consist with the change of 20E titre. Tissues expression analysis revealed OfurGrx transcript expressed abundantly in the head and fat body, compared with that in the intestine, skin and muscle.（4） The OfurGrx expression analysis after 20E challange was studied by using Real- time PCR. The results showed that methoprene （JH analog） had weak or even no effect on OfurGrx expression. But 20E significantly induce OfurGrx expression, which arrive high- est expression after 12 h-20E treatment.（5） OfurGrx expression analysis was studied after adverse factor treatment by using Real-time PCR. The results showed that these adverse factors strongly induce the OfurGrx expression.There is no publication concerned with studies above.