OBJECTIVETo clonea a novel gene related to blood glucose regulation.

METHODSContrast rat model of autonomous regulation through jugular vein right atrium intubation of high concentration of glucose, the control rats were injected with 0.9% NaCl and skeletal muscle was separated. The differentially expressed fragments were identified by differential display technology (DDRT-PCR). After slot blot and Northern blot analysis, we excluded the artificial positive fragments and got the true EST (expression sequence tag) differentially expressed. These positive EST were used as probes to screen cDNA library of rat skeletal muscle, the coding region of full-length gene was cloned to pEGFP and L-6TG cell line was cultured and transiently transfected with the lipofectamine reagent. After 48 h, intact cells were examined with fluorescence microscope.

RESULTSWe got a novel full-length cDNA, named as Fang-1. GenBank Accession No. is AF399873. Using blast software (NCBI), we found Fang-1 is rat homologue of human AK001644. They share 82% identical nucleotides, indicating the family proteins are very conserved. After high concentration of glucose stimulation compared with control rats, the expression of Fang-1 was up-regulated and the expression product of Fang-1 was localized in both cytoplasm and cell nucleus.

CONCLUSIONSA novel gene related to blood glucose regulation was cloned from rat skeletal muscle. The expression product of Fang-1 was localized in both cytoplasm and cell nucleus. The gene can regulate blood glucose level by controlling some mechanisms unknown now with down-regulation expression.