Glucokinase(GK;E.C.2.7.1.1),belonging to hexokinase family,is the liver enzyme that converts glucose into glucose-6-phosphate.It is an important first step in glycogenesis.Here we report a cDNA encoding Erythroculter ...Glucokinase(GK;E.C.2.7.1.1),belonging to hexokinase family,is the liver enzyme that converts glucose into glucose-6-phosphate.It is an important first step in glycogenesis.Here we report a cDNA encoding Erythroculter ilishaeformis GK using RT-PCR and RACE.The cDNA was 2 054 bp with 63 bp 5′UTR,563 bp 3′UTR[excluding poly(A)] and 1 428 bp ORF,which encodes 476 amino acids and has a predicted mol wt of 53.74 KDa.The high similarity between sequences of GK in E.ilishaeformis and GK sequences from other vertebrates confirms that this sequence corresponds to a functional enzyme;The E.ilishaeformis GK appears to possess the ATP-binding site and glucose-binding site,respectively.The deduced amino acid sequence of E.ilishaeformis GK shares 96%,87%,86% sequence identity with common carp,rainbow trout,gilthead seabream,respectively,shares79%,80% identity with clawed frog and human,respectively.For phylogenetic analysis the deduced amino acid sequence of E.ilishaeformis GK,together with GK sequences reported for 12 eukaryotic GK amino acid sequences,were aligned by Clustal W,version 1.6 and the PAUP algorithm was used to produce an unrooted tree.The tree shows E.ilishaeformis GK gene has closely relatedness with other three fish species,and highly homologous to mammalian GK.展开更多
文摘Glucokinase(GK;E.C.2.7.1.1),belonging to hexokinase family,is the liver enzyme that converts glucose into glucose-6-phosphate.It is an important first step in glycogenesis.Here we report a cDNA encoding Erythroculter ilishaeformis GK using RT-PCR and RACE.The cDNA was 2 054 bp with 63 bp 5′UTR,563 bp 3′UTR[excluding poly(A)] and 1 428 bp ORF,which encodes 476 amino acids and has a predicted mol wt of 53.74 KDa.The high similarity between sequences of GK in E.ilishaeformis and GK sequences from other vertebrates confirms that this sequence corresponds to a functional enzyme;The E.ilishaeformis GK appears to possess the ATP-binding site and glucose-binding site,respectively.The deduced amino acid sequence of E.ilishaeformis GK shares 96%,87%,86% sequence identity with common carp,rainbow trout,gilthead seabream,respectively,shares79%,80% identity with clawed frog and human,respectively.For phylogenetic analysis the deduced amino acid sequence of E.ilishaeformis GK,together with GK sequences reported for 12 eukaryotic GK amino acid sequences,were aligned by Clustal W,version 1.6 and the PAUP algorithm was used to produce an unrooted tree.The tree shows E.ilishaeformis GK gene has closely relatedness with other three fish species,and highly homologous to mammalian GK.
