Glucose is the main energy source for mammalian cells and its absorption is co-mediated by two different families of glucose transporters, sodium/glucose co-transporters (SGLTs) and facilitative glucose transporters (GLUTs). Here, we report the cloning and tissue distribution of porcine GLUT2. The GLUT2 was cloned by RACE and its cDNA was 2,051 bp long (GenBank accession no. EF140874). An AAATAA consensus sequence at nucleotide positions 1936-1941 was located upstream of the poly (A) + tail. Open reading frame analysis suggested that porcine GLUT2 contained 524 amino acids, with molecular weight of 57 kDa. The amino acid sequence of porcine GLUT2 was 87% and 79.4% identical with human and mouse GLUT2, respectively. GLUT2 mRNA was detected at highest level in porcine liver, at moderate levels in the small intestine and kidney, and at low levels in the brain, lung, muscle and heart. In the small intestine, the highest level was in the jejunum. In conclusion, the mRNA expression of GLUT2 was not only differentially regulated by age, but also differentially distributed along the small intestine of piglets, which may be related to availability of different intestinal luminal substrate concentrations resulting from different food sources and digestibility.
Cationic amino acid transporter b 0,+ AT (HGMW-approved gene symbol SLC7A9, solute carrier family 7, member 9) plays a crucial role in amino acid nutrition. In the present study, we describe the cloning and sequencing of porcine b 0,+ AT. Based on the sequence of porcine b 0,+ AT deposited in the NCBI (National Center for Biotechnological Information), we identified a putative porcine homologue. Using rapid amplification of cDNA ends (RACE), the full-length cDNA encoding porcine b 0,+ AT was isolated. The porcine b 0,+ AT cDNA was 1,680 bp long, encoding a 487 amino acid trans-membrane protein. The predicted amino acid sequence was found to have 88.9% and 87.1% identity with human and mouse b 0,+ AT, respectively. Real-time RT-PCR indicated porcine b 0,+ AT transcripts expressed in heart, kidney, muscle and small intestine. The small intestine had the highest b 0,+ AT mRNA abundance while the muscle had the lowest (p<0.05). Along the longitudinal axis, the ileum had the highest b 0,+ AT mRNA abundance while the colon had the lowest (p<0.05). The b 0,+ AT mRNA level was highest on day 7 and 90 in the duodenum (p<0.05). It increased from day 1 to day 26 in the jejunum (p>0.05) and had the highest abundance on day 60 (p<0.05). There was, however, no difference between day 1, 7, 26, 30, 90 and 150 (p>0.05). The strongest b 0,+ AT expression appeared on day 7 in the ileum before weaning, and then decreased till day 30 but rose gradually again from day 60 to 150 (p<0.05).
In this study, we cloned, sequenced and characterized porcine y+L Amino Acid Transporter-1 (y+LAT1). By screening a translated EST database with the protein sequence of the human y + LAT1 and by using rapid amplification of cDNA ends (RACE), the full-length cDNA encoding porcine y + LAT1 was isolated from porcine intestine RNA. It was 2,111 bp long, encoding a 511 amino acid trans-membrane glycoprotein composed of 12 transmembrane domains. The predicted amino acid sequence was found to be 91%, 90%, 87% and 87% identical to those of cattle, human, mouse and rat y + LAT1 respectively. Real-time RT-PCR results indicated that the small intestine had the highest y + LAT1 mRNA abundance and the lung had the lowest y + LAT1 mRNA abundance. Baby hamster kidney (BHK) cells transfected with green fluorescent protein (GFP) tagged porcine y + LAT1 cDNA indicated that the cellular localization of the gene product in BHK was on the plasma membrane.
The goal of this study was to elucidate the expression and segmental distribution of the glomerular cationic amino acid metabolism transporter-2 (CAT-2) and thus to improve our understanding of porcine cationic amino acid transporters and amino acid absorption. Porcine CAT-2 was cloned, sequenced and characterized. The predicted amino acid sequence of porcine CAT-2 shared 86.1% and 92.1% identity with human and mouse CAT-2A, respectively. The tissue distribution patterns and ontogenic changes of CAT-2 mRNAs were determined by real-time Q-PCR. The results showed that porcine CAT-2 was highly expressed in the heart and intestinal tract (duodenum, ileum and jejunum). In addition, the mRNA of CAT-2 was found in liver, lung, kidney, brain and muscle. Within the intestinal tract, CAT-2 mRNA was most abundant in the ileum and rarely expressed in the duodenum. In the duodenum, the levels of CAT-2 mRNA reached their peak on day 7 (p<0.05) while in the jejunum, levels were low on day 1 and 7 and increased rapidly after day 26 before peaking on days 30 and 60 (p<0.05). The levels then dramatically decreased by day 90 (p<0.05). In the ileum, levels achieved their maximum on day 30 and then decreased significantly on day 60 (p<0.05).
Molecular cloning, tissue distribution and developmental regulation of porcine GLUT2 (glucose transporter 2) mRNA were conducted in small intestine. GLUT2 cDNA is 2051 bp long, the predicted amino acid sequence revealed that it is highly conserved among vertebrate species. Animal tissue samples were collected from 35 pigs at different developmental stages: sucking (day 1, 7 and 26), postweaning (day 30) and growing (day 60, 90 and 150). The results showed that GLUT2 mRNA was detected at highest level in porcine liver, at moderate levels in the small intestine, kidney, and at low levels in the brain, lung, muscle and heart. In the small intestine, the highest level is in the jejunum. After weaning, the three small intestinal sements took on a similar pattern, GLUT2 mRNA increased through post‐weaning (d30, 2 days post‐weaning) and peaked in growing period (d90), and then decreased in finishing period (d150). In conclusion, the mRNA expression of GLUT2 was not only differentially regulated by age but also differentially distributed along the small intestine of piglets at different stages, which may be related to intestinal luminal substrate concentrations. This study was supported by the National Basic Research Program of China (Project No: 2004CB117501) and National Natural Science Foundation of China (Project No: 30671529).
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