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ABSTRACT: MicroRNAs (miRNAs) are small ∼22 nucleotide regulatory RNAs that regulate the stability and translation of cognate messenger RNAs(mRNAs). MicroRNAs participate in the regulation of adipogenesis and identification of the full repertoire of MicroRNAs expressed in adipose tisse is likely to improve our understanding of adipose tissue growth and development significantly. In the present study, it is found that miR-224-5p abundance decreases first and then increases during adipogenesis of 3T3-L1 cells. And early growth response 2 (EGR2) and Acyl-CoA synthetase long-chain family member 4 (ACSL4) are direct targets of miR-224-5p. Further studies in mouse 3T3-L1 cell-line shows that miR-224-5p is a novel negative regulator of adipocyte differentiation through post-transcriptional regulation of early growth response 2 during early adipogenesis. Furthermore, miR-224-5p could regulate fatty acid metabolism through Acyl-CoA synthetase long-chain family member 4 at terminal differentiation. It indicates that miR-224 plays different roles on different stages of adipogenesis.
The international journal of biochemistry & cell biology 05/2013; · 4.89 Impact Factor
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ABSTRACT: Lysine-specific demethylase 1 (LSD1) functioned as a demethyl methylase gene, underlying a wide range of biological processes, including cancer, cell apoptosis, differentiation, and development. To further understand the functions of the porcine LSD1 gene, we first obtained cDNA sequence of porcine LSD1 gene, using in silico cloning method. We further found that the porcine LSD1 gene has two transcripts, in which cDNA sequences are 2,716 and 2,656 bp, ORF are 2,622 and 2,562 bp, respectively. Then, RT-PCR analysis showed that the LSD1 gene is expressed in various tissues and relatively higher in the tissues of ovary, kidney, and spleen. Besides, the LSD1 gene was expressed higher in the growth nonage and peaked at 3 days in muscle tissue. Meanwhile, the expression of two transcript variants of LSD1 gene presented the same change trend. Besides, the level of DNA methylation was approximately fourfold higher in a 3-day muscle than in an old pig (180 days), significantly positive related to the gene expression of LSD1 (R = 0.9362, P < 0.05), and declined with growing age. Cloning, expression pattern, and analysis with genome DNA methylation of porcine LSD1 gene laid a foundation to clarify the molecular mechanisms of porcine growth and development and also for further work on animal breeding.
Applied biochemistry and biotechnology 11/2012; · 1.94 Impact Factor
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Qi Xiong,
Jin Chai,
Changyan Deng,
Siwen Jiang,
Yang Liu,
Tao Huang,
Xiaojun Suo,
Nian Zhang,
Xiaofeng Li,
Qianping Yang,
Mingxin Chen, Rong Zheng
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ABSTRACT: Skeletal muscle and kidney-enriched inositol phosphatase (SKIP) was identified as a 5'-inositol phosphatase that hydrolyzes phosphatidylinositol (3,4,5)-triphosphate (PI(3,4,5)P3) to PI(3,4)P2 and negatively regulates insulin-induced phosphatidylinositol 3-kinase signaling in skeletal muscle. In this study, two new single nucleotide polymorphisms (SNPs) in porcine SKIP introns 1 and 6 were detected. The C1092T locus in intron 1 showed significant associations with some meat traits, whereas the A17G locus in intron 6 showed significant associations with some carcass traits. Expression analysis showed that porcine SKIP is upregulated at d 65 of gestation and Meishan fetuses have higher and prolonged expression of SKIP compared to Large White at d 100 of gestation. Ectopic expression of porcine SKIP decreased insulin-induced cell proliferation and promoted serum starvation-induced cell cycle arrest in G0/G1 phase in C2C12. Our results suggest that SKIP plays a negative regulatory role in skeletal muscle development partly by preventing cell proliferation.
Meat Science 05/2012; 92(4):490-7. · 2.28 Impact Factor
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ABSTRACT: The domestic pig (Sus scrofa), an important species in animal production industry, is a right model for studying adipogenesis and fat deposition. In order to expand the repertoire of porcine miRNAs and further explore potential regulatory miRNAs which have influence on adipogenesis, high-throughput Solexa sequencing approach was adopted to identify miRNAs in backfat of Large White (lean type pig) and Meishan pigs (Chinese indigenous fatty pig). We identified 215 unique miRNAs comprising 75 known pre-miRNAs, of which 49 miRNA*s were first identified in our study, 73 miRNAs were overlapped in both libraries, and 140 were novelly predicted miRNAs, and 215 unique miRNAs were collectively corresponding to 235 independent genomic loci. Furthermore, we analyzed the sequence variations, seed edits and phylogenetic development of the miRNAs. 17 miRNAs were widely conserved from vertebrates to invertebrates, suggesting that these miRNAs may serve as potential evolutional biomarkers. 9 conserved miRNAs with significantly differential expressions were determined. The expression of miR-215, miR-135, miR-224 and miR-146b was higher in Large White pigs, opposite to the patterns shown by miR-1a, miR-133a, miR-122, miR-204 and miR-183. Almost all novel miRNAs could be considered pig-specific except ssc-miR-1343, miR-2320, miR-2326, miR-2411 and miR-2483 which had homologs in Bos taurus, among which ssc-miR-1343, miR-2320, miR-2411 and miR-2483 were validated in backfat tissue by stem-loop qPCR. Our results displayed a high level of concordance between the qPCR and Solexa sequencing method in 9 of 10 miRNAs comparisons except for miR-1a. Moreover, we found 2 miRNAs, miR-135 and miR-183, may exert impacts on porcine backfat development through WNT signaling pathway. In conclusion, our research develops porcine miRNAs and should be beneficial to study the adipogenesis and fat deposition of different pig breeds based on miRNAs.
PLoS ONE 01/2012; 7(2):e31426. · 4.09 Impact Factor
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Qi Xiong,
Jin Chai,
Changyan Deng,
Siwen Jiang,
Xiaofeng Li,
Xiaojun Suo,
Nian Zhang,
Qianping Yang,
Yang Liu, Rong Zheng,
Mingxin Chen
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ABSTRACT: Src homology 2-containing inositol 5-phosphatase 2 (SHIP2) has been identified as 5'-inositol phosphatase that hydrolyzes PI(3,4,5)P(3) to PI(3,4)P(2), which negatively regulates insulin-induced Akt signaling in skeletal muscle. In this study, we obtained a 3,795-bp mRNA sequence of porcine SHIP2 that included the full coding region for a protein of 1,264 amino acids. With the use of comparative mapping, we mapped this gene to SSC9 p23-24, where many QTLs affect average backfat thickness, average daily weight gain (birth-10 weeks), adipocyte number, belly fat area, and mid-back fat traits. As a candidate gene for carcass traits, a novel single nucleotide polymorphism in intron 21 (A > G) was detected by PCR-RFLP. The results showed that the AA genotype had higher skin percentage, shoulder fat thickness, and m. longissimus dorsi width, but lower m. longissimus dorsi height compared with the genotype GG (P < 0.05), and that allele G appeared to be associated with an increase in the growth trait. SHIP2 was expressed abundantly in skeletal muscle tissue and was transcriptionally decreased during the proliferative phase, but increased in the intermediate stages of muscle differentiation. Analysis of the porcine SHIP2 promoter sequence demonstrated that the E2F element is involved in downregulating SHIP2 mRNA expression in proliferating myoblasts. Using RNAi, we found that the MyoD transcription factor played a role in upregulating SHIP2 expression in differentiating myotubes. In summary, we suggest that SHIP2 might play a role in the regulation of skeletal muscle development in pigs.
Molecular and Cellular Biochemistry 09/2011; 360(1-2):225-33. · 2.06 Impact Factor
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ABSTRACT: The temporal and spatial distribution of the genus Bacillus and Clostridium histolyticum group in swine manure composting was determined by fluorescent in situ hybridization using fluorescently labeled 16S rRNA-targeted oligonucleotide probes LGC353b and Chis150, respectively. The temporal distribution of total bacteria, Bacillus and C. histolyticum, detected in each layer of the composting pile was noticeable in that the number of them detected at the high-temperature stage was higher than that of the cooling stage. The number detected at the cooling stage was higher than that of the temperature-rising stage. The number of the total bacteria distributed in three locations achieved balance at the stage of cooling. The spatial distribution of the genus Bacillus cells was that the number and the relative abundance of Bacillus cells detected in the middle layer of composting pile were the lowest at each stage of composting. However, the minimum value of the relative abundance exceeded 8%. Compared with Bacillus spp., the C. histolyticum group displayed higher relative abundance in the same layer at different stages of composting except in the top layer at the stage of high temperature. However, the characteristic of the spatial distribution was not noticeable. The detected limits of the genus Bacillus and C. histolyticum group were both found to be the high cell density of 10(6) cells g(-1) (wet weight). These results indicated that the genus Bacillus and C. histolyticum group were the predominant bacteria in the swine manure composting process and may play important role in this complex environment.
Applied Microbiology and Biotechnology 09/2011; 93(6):2625-32. · 3.42 Impact Factor
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ABSTRACT: In order to obtain the diversity and temporal-spatial distribution of Bacillus community during the swine manure composting, we utilized traditional culture methods and the modern molecular biology techniques of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and -denaturing gradient gel electrophoresis (PCR-DGGE). Bacillus species were firstly isolated from the composting. Based on temperature changes, the temporal-spatial characteristics of total culturable Bacillus were remarkable that the number of the culturable Bacillus detected at the high-temperature stage was the highest in each layer of the pile and that detected in the middle layer was the lowest at each stage of composting respectively. The diversity of cultivated Bacillus species isolated from different composting stages was low. A total of 540 isolates were classified by the RFLP method and partial 16S rDNA sequences. They affiliated to eight species including Bacillus subtilis, Bacillus cereus, Bacillus thuringiensis, Bacillus anthracis, Bacillus megaterium, Bacillus licheniformis, Bacillus pumilus, and Bacillus circulans. The predominant species was B. subtilis, and the diversity of culturable Bacillus isolated in the middle-level samples at temperature rising and cooling stages was the highest. The DGGE profile and clone library analysis revealed that the temporal-spatial distribution of Bacillus community was not obvious, species belonging to the Bacillus were dominant (67%) with unculturable bacteria and B. cereus was the second major culturable Bacillus species. This study indicated that a combination of culture and culture-independent approaches could be very useful for monitoring the diversity and temporal-spatial distribution of Bacillus community during the composting process.
Applied Microbiology and Biotechnology 06/2011; 93(1):411-21. · 3.42 Impact Factor
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ABSTRACT: Imprinted genes play important roles in mammalian growth, development and behavior. In this study, we obtained 1568 bp mRNA sequence of porcine DIO3 (deiodinase, iodothyronine, type III), and also identified its imprinting status during porcine fetal development. The complete open reading frame (ORF) encoding 278 amino acids. The porcine DIO3 mRNA was expressed predominantly in backfat, mildly in liver, uterus, kidney, heart, small intestine, muscle and stomach, and almost absent in spleen and lung. A single nucleotide polymorphism in exon (A/C (687)) was used to investigate the allele frequencies in different pig breeds and the imprinting status in porcine embryonic tissues. The results indicate that DIO3 was imprinted in all the tested tissues. Statistical analysis showed the DIO3 gene polymorphism was significantly associated with almost all the fat deposition and carcass traits, including lean meat percentage (LMP), fat meat percentage (FMP), ratio of lean to fat (RLF), shoulder fat thickness (SFT), sixth-seventh rib fat thickness (RFT), buttock fat thickness (BFT), loin eye area (LEA), and intramuscular fat (IMF).
Molecular Biology Reports 06/2011; 39(3):2329-35. · 2.93 Impact Factor
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ABSTRACT: Imprinted genes play an essential role in the regulation of fetal growth, development and function of the placenta, however only a limited number of imprinted genes have been studied in swine. In this study, we cloned and characterized porcine MAGEL2 (melanoma antigen-like gene 2), and also identified its imprinting status during porcine fetal development. The complete open reading frame (ORF) encoding 1,193 amino acids was isolated and two single nucleotide polymorphisms (SNPs) (g.2592A>C and g.3277T>C) in the coding region were identified. The reciprocal Yorkshire×Meishan F1 hybrid model and the RT-PCR/RFLP method were used to detect the imprinting status of porcine MAGEL2 gene at two developmental stages of day 30 and 65 of gestation. Imprinting analysis showed that porcine MAGEL2 was paternally expressed in day 65 fetal tissues, including heart, liver, spleen, lung, kidney, stomach, small intestine, skeletal muscle, brain and placenta. Interestingly, we observed an imprinting variance of MAGEL2 gene in 30 dpc fetuses produced by the cross of Yorkshire boar×Meishan sow, in which seven heterozygous fetuses were monoallelically expressed from the paternal allele but two were biallelically expressed from both the paternal and maternal alleles. Association analysis in a Yorkshire×Meishan F2 resource population showed that the mutation of g.2592A>C was significantly associated with dressed carcass percentage (P<0.05) and buttock fat thickness (P<0.05). Our results suggest that MAGEL2, as a novel imprinted gene in pig, might be a candidate gene affecting carcass traits and could provide important information for the functional study of imprinted genes during porcine development.
Molecular Biology Reports 06/2011; 39(1):147-55. · 2.93 Impact Factor
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ABSTRACT: Obese and lean pig breeds show obvious differences in adipose metabolism/fat deposition; however, the molecular mechanism underlying phenotype variation remains unknown. In order to understand it, we analyzed the differences of gene expression in backfat between Meishan (a typical Chinese indigenous obese breed) and Large White (a lean Western breed) pigs. Here, we cloned porcine β subunit of IDH3 (IDH3B) and 2447 bp 5'-flanking sequence of this gene, and determined the genomic structure. Porcine IDH3B contains three isoforms, IDH3B ( 1 ), IDH3B ( 2 ) and IDH3B ( 3 ). Real-time RT-PCR revealed that these three isoforms were prevalently up-regulated in backfat of western commercial pigs, Large White, Landrace and Duroc, compared with Chinese indigenous breeds, Meishan and Tongcheng pigs. A 304 bp insertion/deletion variant was found in the 5'-flanking region. Dual-luciferase reporter assays showed that in vitro the promoter of IDH3B gene with the insertion had higher luciferase activity as compared with the wild type. Three genotypes AA, AB and BB, due to this insertion, were detected, and the frequency of allele A was dominant in western commercial pigs, whereas allele B predominated in Chinese indigenous breeds. IDH3B mRNA expression in Meishan pigs was more abundant with genotype AA than with genotype AB or BB, as in Large White pigs. In addition, the polymorphism was detected in 317 pigs of a Large White × Meishan F2 resource population. Association analysis showed that pigs with genotype AA possessed higher backfat thickness at buttocks than those with genotype AB (P < 0.05) or BB. These data suggested that the 304 bp insertion mutation in promoter region increased the expression of porcine IDH3β transcripts and this mutation might be a candidate marker for marker assistant selection in swine breeding.
Molecular Biology Reports 05/2011; 39(2):1419-26. · 2.93 Impact Factor
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ABSTRACT: The myocyte enhancer factor 2C (MEF2C) is a member of the MEF2 family of transcription factors, involved in skeletal muscle development. In this study we report the cDNA sequence and isolate the 5' upstream region of the mef2c gene from porcine genomic DNA using PCR-based GenomeWalker. The open reading frame of porcine mef2c cDNA covers 1,392 bases, encoding 464 amino acids, which show 94% identity with human MEF2C at the level of the primary protein structure. Annear the C terminus of mef2c, a 96-nt sequence appear to represent alternatively spliced transcripts was present in some cDNAs and absent in the other. No typical TATA, GC box or CAAT box binding site was found in porcine mef2c 5' upstream region, whereas some potential binding sites for MyoD (E-box), MEF2 and MBF1 were present in the proximal upstream region. Transfection of the mef2c 5' upstream region with EGFP into cos7 cells demonstrated that the region from -162 to +115 bp immediately 5' of the exon 1 was sufficient to direct strong EGFP protein expression. Co-transfection assays demonstrated that MBF1 bound the mef2c promoter and inhibited mef2c expression. These results may be useful for elucidating the regulation mechanisms of mef2c, which interacts with other factors to regulate target genes.
Molecular Biology Reports 12/2010; 38(7):4723-30. · 2.93 Impact Factor
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ABSTRACT: Ladybird-like genes were recently identified in mammals. The first member characterized, Lbx1, is expressed in developing skeletal muscle and the nervous system. However, little is known about the porcine Lbx1 gene. In the present study, we cloned and characterized Lbx1 from porcine muscle. RT-PCR analyses showed that Lbx1 was highly expressed in porcine skeletal muscle tissues. And we provide the first evidence that Lbx1 has a certain regulated expression pattern during the postnatal period of the porcine skeletal muscle development. Lbx1 gene expressed at higher levels in biceps femoris muscles compared with masseter, semitendinosus and longissimus dorsi muscles in Meishan pigs. Phylogenetic tree was constructed by aligning the amino acid sequences of different species. Moreover, single nucleotide polymorphism (SNP) scanning in the Lbx1 genomic fragment identified two mutations, g.752A>G and g.-1559C>G. Association analysis in our experimental pig populations showed that the mutation of g.752A>G was significantly associated with loin muscle area (P<0.05) and internal fat rate (P<0.05). Our results suggest that the Lbx1 gene might be a candidate gene of carcass traits and provide useful information for further studies on its roles in porcine skeletal muscle.
Molecular Biology Reports 11/2010; 38(6):3983-91. · 2.93 Impact Factor
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Lifan Luo,
Lianzhi Ye,
Gang Liu,
Guochao Shao, Rong Zheng,
Zhuqing Ren,
Bo Zuo,
Dequan Xu,
Minggang Lei,
Siwen Jiang,
Changyan Deng,
Yuanzhu Xiong,
Fenge Li
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ABSTRACT: Background
MicroRNAs (miRNAs) are short non-coding RNA molecules which are proved to be involved in mammalian spermatogenesis. Their expression and function in the porcine germ cells are not fully understood.
Methodology
We employed a miRNA microarray containing 1260 unique miRNA probes to evaluate the miRNA expression patterns between sexually immature (60-day) and mature (180-day) pig testes. One hundred and twenty nine miRNAs representing 164 reporter miRNAs were expressed differently (p
PLoS ONE 08/2010; 5(8). · 4.09 Impact Factor
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Jin Chai, Qi Xiong, Pengpeng Zhang, Rong Zheng,
Jian Peng, Siwen Jiang
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ABSTRACT: Glucocorticoid (GC) are stress hormones, whose cytotoxicity has been shown in various cells. The imbalance of calcium homeostasis is believed to be associated with the dexamethasone (DEX, a synthetic GC)-induced apoptosis. Here we show that in C2C12 myoblasts, DEX markedly up-regulated the expression of inositol 1,4,5-triphosphate receptor 1 (IP3R1) and down-regulated the expression of SERCA1 (sarcoendoplasmic reticulum Ca2+-ATPase 1), leading to calcium overload. Furthermore, the imbalance of calcium homeostasis increased the level of BAX, decreased the level of Bcl-2, induced cytochrome c release and activated caspase-3, leading to intranucleosomal DNA fragmentation and plasma membrane damage, eventually resulting in cell apoptosis. Taken together, by using C2C12 myoblasts as a model system, we demonstrated a novel mechanism for stress hormone-induced apoptosis: it is dependent on the induction of intracellular calcium overload via the alterations of IP3R1 and SERCA1 expressions.
General and Comparative Endocrinology - GEN COMP ENDOCRINOL. 01/2010; 166(2):241-249.
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Lifan Luo,
Lianzhi Ye,
Gang Liu,
Guochao Shao, Rong Zheng,
Zhuqing Ren,
Bo Zuo,
Dequan Xu,
Minggang Lei,
Siwen Jiang,
Changyan Deng,
Yuanzhu Xiong,
Fenge Li
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ABSTRACT: MicroRNAs (miRNAs) are short non-coding RNA molecules which are proved to be involved in mammalian spermatogenesis. Their expression and function in the porcine germ cells are not fully understood.
We employed a miRNA microarray containing 1260 unique miRNA probes to evaluate the miRNA expression patterns between sexually immature (60-day) and mature (180-day) pig testes. One hundred and twenty nine miRNAs representing 164 reporter miRNAs were expressed differently (p<0.1). Fifty one miRNAs were significantly up-regulated and 78 miRNAs were down-regulated in mature testes. Nine of these differentially expressed miRNAs were validated using quantitative RT-PCR assay. Totally 15,919 putative miRNA-target sites were detected by using RNA22 method to align 445 NCBI pig cDNA sequences with these 129 differentially expressed miRNAs, and seven putative target genes involved in spermatogenesis including DAZL, RNF4 gene were simply confirmed by quantitative RT-PCR.
Overall, the results of this study indicated specific miRNAs expression in porcine testes and suggested that miRNAs had a role in regulating spermatogenesis.
PLoS ONE 01/2010; 5(8):e11744. · 4.09 Impact Factor
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ABSTRACT: Glucocorticoid (GC) are stress hormones, whose cytotoxicity has been shown in various cells. The imbalance of calcium homeostasis is believed to be associated with the dexamethasone (DEX, a synthetic GC)-induced apoptosis. Here we show that in C2C12 myoblasts, DEX markedly up-regulated the expression of inositol 1,4,5-triphosphate receptor 1 (IP3R1) and down-regulated the expression of SERCA1 (sarcoendoplasmic reticulum Ca(2+)-ATPase 1), leading to calcium overload. Furthermore, the imbalance of calcium homeostasis increased the level of BAX, decreased the level of Bcl-2, induced cytochrome c release and activated caspase-3, leading to intranucleosomal DNA fragmentation and plasma membrane damage, eventually resulting in cell apoptosis. Taken together, by using C2C12 myoblasts as a model system, we demonstrated a novel mechanism for stress hormone-induced apoptosis: it is dependent on the induction of intracellular calcium overload via the alterations of IP3R1 and SERCA1 expressions.
General and Comparative Endocrinology 09/2009; 166(2):241-9. · 3.27 Impact Factor
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ABSTRACT: Apolipoprotein M (APOM), a novel apolipoprotein presented mostly in high-density lipoprotein (HDL) in plasma, is involved in lipid and lipoprotein metabolism. Through comparative mapping, we have mapped this gene to SSC7 p1.1 in which many QTLs affecting fat deposition traits have been reported. As a candidate gene for fat deposition traits, in this study, we obtained the 742-bp mRNA sequence of porcine APOM including the full coding region and encoding a protein of 188 amino acids. The sequence was deposited into the GenBank under the accession no. DQ329240. Semi-quantitative RT-PCR results showed that the porcine APOM gene is expressed predominantly in liver and kidney tissue. The genomic sequence of this gene which contains six exons and five introns, is 3,621 bp in length (DQ272488). Bioinformatic analysis of the 5' regulatory region has revealed that classical TATA-box element and species conserved Hepatocyte nuclear factor-1a (HNF-1alpha) biding site were represented in this region. A G2289C single nucleotide polymorphism (SNP) in the intron 2 of porcine APOM gene detected as an Eco130I PCR-restriction fragment length polymorphism (PCR-RFLP) showed allele frequency differences among three purebreds. Association of the genotypes with fat deposition traits showed that different genotypes of porcine APOM gene were significantly associated with leaf fat weight (P < 0.05), backfat thickness at shoulder (P < 0.05), backfat thickness at thorax-waist (P < 0.05), backfat thickness at buttock (P < 0.01) and average backfat thickness over shoulder, thorax-waist and buttock (P < 0.01).
Molecular Biology Reports 04/2009; 37(3):1363-71. · 2.93 Impact Factor
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ABSTRACT: PI(3,4,5)P(3) produced by the activated PI3-kinase is a key lipid second messenger in cell signaling downstream of insulin. Skeletal muscle and kidney-enriched inositol phosphatase (SKIP) identified as a 5'-inositol phosphatase that hydrolyzes PI(3,4,5) P(3) to PI(3,4)P(2), negatively regulates the insulin-induced glycogen synthesis in skeletal muscle. However the mechanism by which this occurs remains unclear. To elucidate the function of SKIP in glycogen synthesis, we employed RNAi techniques to knockdown the SKIP gene in differentiating C2C12 myoblasts. Insulin-induced phosphorylation of Akt (protein kinase B) and GSK-3beta (Glycogen synthase kinase), subsequent dephosphorylation of glycogen synthase and glycogen synthesis were increased by inhibiting the expression of SKIP, whereas the insulin-induced glycogen synthesis was decreased by overexpression of WT-SKIP. Our results suggest that SKIP plays a negative regulatory role in Akt/ GSK-3beta/GS (glycogen synthase) pathway leading to glycogen synthesis in myocytes.
BMB reports 03/2009; 42(2):119-24. · 1.72 Impact Factor
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Min Liu,
Jian Peng,
De-Quan Xu, Rong Zheng,
Feng-E Li,
Jia-Lian Li,
Bo Zuo,
Ming-Gang Lei,
Yuan-Zhu Xiong,
Chang-Yan Deng,
Si-Wen Jiang
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ABSTRACT: MYF5 and MYOD1 belong to the myogenic regulatory factor (MRF) gene family. They code for the basic helix-loop-helix transcription factors that play key regulatory roles in the initiation and development of skeletal muscle and the maintenance of its phenotype. In this work three single nucleotide polymorphisms (SNPs) in porcine MYF5 and one in porcine MYOD1 were detected in three pig breeds (Large White, Landrace, and Meishan) by means of a PCR-RFLP protocol. Analysis of the association of meat quality traits with the four polymorphisms in a series of three Large White x Meishan F2 populations, totaling 399 pigs, found: (1) MYF5 exon 1 Hsp92II polymorphism causing a Met --> Leu substitution was associated with intramuscular fat content (P = 0.04) and water moisture content (P = 0.0001) in the longissimus dorsi; (2) MYF5 exon 2 MspI polymorphism and an intron 1 HaeIII polymorphism, which were completely linked, were significantly associated with longissimus dorsi pH (P < 0.05); (3) MYOD1 intron 1 DdeI polymorphism was not significantly associated with any meat quality traits tested. Among these genetic variants (a novel SNP and three identified SNPs), our data suggested that the novel SNP of the MYF5 gene within exon 1 is valuable for pig breeding.
Biochemical Genetics 09/2008; 46(11-12):720-32. · 0.86 Impact Factor
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Zhu-Qing Ren,
Yan Wang,
Yong-Jie Xu,
Lin-Jie Wang,
Ming-Gang Lei,
Bo Zuo,
Feng-E Li,
De-Quan Xu, Rong Zheng,
Chang-Yan Deng,
Si-Wen Jiang,
Yuan-Zhua Xiong
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ABSTRACT: Abstract
ATP-citrate lyase (ACL), one of the lipogenic enzymes, catalyses the formation of acetyl-coenzyme A (CoA) involved in the synthesis of fatty acid and cholesterol. In pig, very little is known about the ACL gene. In this work, the mRNA differential display technique was used to analyse the differences in gene expression between Meishan and Large White pigs and the F1 hybrids of both direct and reciprocal crosses. Our results show that among the differentially expressed genes ACL is up-regulated in the backfat of the F1 hybrids. After cloning and analysing the fulllength cDNA and the 870 bp 5'-flanking sequence of the porcine ACL gene, a C/T mutation at position -97 bp upstream of the transcription site was detected. Luciferase activity detection showed that this mutation changed the transcriptional activity. In F1 hybrids, the heterozygous genotype CT was more frequent than the homozygous genotypes CC and TT . Real-time PCR analysis showed that in Meishan pigs, ACL mRNA expression was more abundant in individuals with genotype CT than in those with genotype CC or TT or in Large White pigs. These results indicate that the C/T mutation affects ACL mRNA expression, probably via the activator protein 2.
Genetics Selection Evolution. 01/2008;
