A full-length cDNA coding glucokinase (GK) was cloned from Pengze crucian carp (Carassius auratus var. Pengze) by RT-PCR and rapid amplification of cDNA ends (RACE) methods. The cDNA sequence obtained is of 2 050 bp length with a 1 431 bp open reading frame (ORF) encoding 476 amino acids. The GK protein has a calculated molecular weight of 53.78 ku and isolectric point of 5.14. The main domains of GK, such as ATP-binding domain, glucose-binding amino acids, hexokinases signature, N-linked glycosylation sites, cell attachment sequence and glycosaminoglycan attachment site for the Pengze crucian carp are basically conservative compared with other vertebrates. The amino acid sequence has a high similarity to GK of other species, and the percent identity compared with common carp, human and rat are 98.1%, 79.8% and 79.1%, respectively. Tissue distribution of GK mRNA in brain, white muscle, spleen, mesenteric adipose tissue and liver of Pengze crucian carp was analyzed by semi-quantitative RT-PCR method using β-actin gene as internal control. The result showed that the expression level of GK mRNA in liver was significantly higher than that in spleen, mesenteric adipose tissue and brain (P<0.05). GK mRNA did not be detected in white muscle.［Chinese Journal of Animal Nutrition, 2011, 23（7）：1167 -1175］
CHENG Hanliang,JI Nanjing,PENG Yongxing,SHEN Xin,WU Chenchen,XU Jianhe,DONG Zhiguo
. Cloning and Expression Analysis of a Full-length cDNA Encoding Glucokinase from Pengze Crucian Carp (Carassius auratus var. Pengze)[J]. Chinese Journal of Animal Nutrition, 2011
, 23(07)
: 1167
-1175
.
DOI: 10.3969/j.issn.1006-267x.2011.07.015
［1］KAWAI S, MUKAI T, MORI S, et al. Hypothesis: structures, evolution, and ancestor of glucose kinases in the hexokinase family[J]. Journal of Bioscience and Bioengineering, 2005, 99(4):320-330.
[2]MAHALINGAM B, CUESTA-MUNOZ A, DAVIS E A, et al. Structural model of human glucokinase in complex with glucose and ATP: implications for the mutants that cause hypo- and hyperglycemia[J]. Diabetes, 1999, 48(9):1698-1705.
[3]EGEA M, METON I, CORDOBA M, et al. Role of Sp1 and SREBP-1a in the insulin-mediated regulation of glucokinase transcription in the liver of gilthead sea bream (Sparus aurata)[J]. General and Comparative Endocrinology, 2008, 155(2):359-367.
[4]STOFFEL M, FROGUEL P, TAKEDA J, et al. Human glucokinase gene: isolation, characterization, and identification of two missense mutations linked to early-onset non-insulin-dependent (type 2) diabetes mellitus[J]. Proceedings of the National Academy of Sciences of the United States of America, 1992, 89(16):7698-7702.
[5]HEMRE G I, MOMMSEN T P, KROGDAHL U. Carbohydrates in fish nutrition: effects on growth, glucose metabolism and hepatic enzymes[J]. Aquaculture Nutrition, 2002, 8(3):175-194.
[6]STONE D A J. Dietary carbohydrate utilization by fish[J]. Reviews in Fisheries Science, 2003, 11:337-370.
[7]CASERAS A, METON I, FERNANDEZ F, et al. Glucokinase gene expression is nutritionally regulated in liver of gilthead sea bream (Sparus aurata)[J]. Biochimica et Biophysica Acta, 2000, 1493(1/2):135-141.
[8]MOON T W. Glucose intolerance in teleost fish: fact or fiction?[J]. Comparative Biochemistry and Physiology Part B: Biochemistry & Molecular Biology, 2001, 129(2/3):2-3.
[9]程汉良,夏德全,吴婷婷.鱼类脂类代谢调控与脂肪肝[J].动物营养学报,2006,18(4):294-298.
[10]BLIN C, PANSERAT S, KAUSHIK S, et al. Partial molecular cloning and tissue distribution of hexokinase Ⅰ cDNA in common carp[J]. Journal of Fish Biology, 2000, 56:1558-1561.
[11]PANSERAT S, BLIN C, MEDALE F, et al. Molecular cloning, tissue distribution and sequence analysis of complete glucokinase cDNAs from gilthead seabream (Sparus aurata), rainbow trout (Oncorhynchus mykiss) and common carp (Cyprinus carpio)[J]. Biochimica et Biophysica Acta, 2000, 1474(1):61-69.
[12]戈贤平,俞菊华,吴婷婷.翘嘴红鮊GK基因的克隆和序列分析[J].水产学报,2006,30(3):410-415.
[13]DIEFFENBACH C W, DVEKSLER G S. PCR primer: a laboratory manual[M]. Plainview, N.Y.: Cold Spring Harbor Laboratory Press, 1995.
[14]KUMAR S, TAMURA K, NEI M. MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment[J]. Briefings in Bioinformatics, 2004, 5(2):150-163.
[15]ANDREONE T L, PRINTZ R L, PILKIS S J, et al. The amino acid sequence of rat liver glucokinase deduced from cloned cDNA[J]. The Journal of Biological Chemistry, 1989, 264(1):363-369.
[16]VEIGA-DA-CUNHA M, COURTOIS S, MICHEL A, et al. Amino acid conservation in animal glucokinases. Identification of residues implicated in the interaction with the regulatory protein[J]. The Journal of Biological Chemistry, 1996, 271(11):6292-6297.
[17]PANSERAT S, FONTAGNE S, BERGOT P, et al. Ontogenesis of hexokinase Ⅰ and hexokinase Ⅳ (glucokinase) gene expressions in common carp (Cyprinus carpio) related to diet[J]. The British Journal of Nutrition, 2001, 85(6):649-651.
[18]SOENGAS J L, POLAKOF S, CHEN X, et al. Glucokinase and hexokinase expression and activities in rainbow trout tissues: changes with food deprivation and refeeding[J]. American Journal of Physiology: Regulatory, Integrative and Comparative Physiology, 2006, 291(3):810-821.
[19]PANSERAT S, CAPILLA E, GUTIERREZ J, et al. Glucokinase is highly induced and glucose-6-phosphatase poorly repressed in liver of rainbow trout (Oncorhynchus mykiss) by a single meal with glucose[J]. Comparative Biochemistry and Physiology Part B: Biochemistry & Molecular Biology, 2001, 128(2):275-283.
[20]CAPILLA E, MEDALE F, NAVARRO I, et al. Muscle insulin binding and plasma levels in relation to liver glucokinase activity, glucose metabolism and dietary carbohydrates in rainbow trout[J]. Regulatory Peptides, 2003, 110(2):123-132.
[21]PANSERAT S, MEDALE F, BLIN C, et al. Hepatic glucokinase is induced by dietary carbohydrates in rainbow trout, gilthead seabream, and common carp[J]. American Journal of Physiology: Regulatory, Integrative and Comparative Physiology, 2000, 278(5):1164-1170.
[22]METON I, CASERAS A, FERNANDEZ F, et al. Molecular cloning of hepatic glucose-6-phosphatase catalytic subunit from gilthead sea bream (Sparus aurata): response of its mRNA levels and glucokinase expression to refeeding and diet composition[J]. Comparative Biochemistry and Physiology Part B: Biochemistry & Molecular Biology, 2004, 138(2):145-153.
[23]FERNANDEZ F, MIQUEL A G, CORDOBA M, et al. Effects of diets with distinct protein-to-carbohydrate ratios on nutrient digestibility, growth performance, body composition and liver intermediary enzyme activities in gilthead sea bream (Sparus aurata L.) fingerlings[J]. Journal of Experimental Marine Biology and Ecology, 2007, 343(1):1.
[24]EGEA M, METON I, BAANANTE I V. Sp1 and Sp3 regulate glucokinase gene transcription in the liver of gilthead sea bream (Sparus aurata)[J]. Journal of Molecular Endocrinology, 2007, 38(4):481-492.
[25]ENES P, PANSERAT S, KAUSHIK S, et al. Hepatic glucokinase and glucose-6-phosphatase responses to dietary glucose and starch in gilthead sea bream (Sparus aurata) juveniles reared at two temperatures[J]. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 2008, 149(1):80-86.
