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PPARD基因在动物脂肪代谢过程中起重要作用，本研究以猪为实验动物，通过RT-PCR方法扩增得到PPARD基因全长编码区序列，在编码区第95位检测到G/A两种核苷酸，序列提交GenBank数据库获得登录号“KP757025和KP757026”。生物信息学分析显示：该基因编码462个氨基酸，蛋白分子量为49.74 kD，等电点为7.53。蛋白质结构分析PPARD存在2个保守域，无跨膜结构，不存在信号肽，存在1个亮氨酸富集的核输出信号，其N端、C端均亲水；亚细胞定位显示该蛋白分泌到细胞周质的概率为69.6%。系统进化分析表明，猪与牛、家犬的亲缘关系较近。利用qPCR检测了PPARD基因mRNA在猪14个组织中的表达量，发现在肝脏、耳朵、脾脏、小肠中表达量较高。本研究为进一步研究猪PPARD基因的表达调控奠定研究基础。 PPARD as the main fat metabolism genes, for studying the regulation and function of PPARD in pigs, PPARD cDNA including the coding sequence was obtained by RT-PCR with the RNA isolated from pigs and submitted to GenBank with the access number. Bioinformatics analysis showed that the PPARD gene encode a protein of 462 amino acids with a predicted molecular weight of 49.74 kD and the isoelectric point of 7.53. Protein structure analysis showed that PPARD protein contains two conserved domain, no transmembrane regions and signal peptide and had a leucine-rich nuclear export signals． Hydrophobicity analysis indicate that the N-terminus and C-terminus are hydrophilic. The probability of secreting into periplasmic is 69.6%. Phylogenetic results indicated that the pig has the closest relationship with cattle and dog. We detected G and A two kinds nucleotides at 95 nucleotide in the coding region of the PPARD gene and three genotypes were found: AA, AB, BB. We tested the PPARD mRNA expression in 14 tissues by qPCR, the result showed that the expression is high in liver, spleen, ear, small intestine. This study provides experimental basis for further study on the expression and regulation of PPARD gene. |
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huojinlong8610: 金币+100, 翻译EPI+1 2015-02-11 15:28:27
huojinlong8610: 金币+100, 翻译EPI+1 2015-02-11 15:28:27
| PPARδ is a nuclear hormone receptor that governs a variety of biological processes, including fatty acid metabolism. For studying the regulation and function of PPARD in pigs, PPARδ cDNA including the coding sequence was obtained by RT-PCR with the RNA isolated from pigs and submitted to GenBank with the access number. Bioinformatics analysis showed that the PPARδ gene encode a protein of 462 amino acids with a predicted molecular weight of 49.74 kD and the isoelectric point of 7.53. Protein structure analysis showed that PPARδ protein contains two conserved domains and a leucine-rich nuclear export signal without transmembrane regions and signal sequence． Hydrophobicity analysis indicate that the N-terminus and C-terminus are hydrophilic. The probability of secreting into periplasmic is 69.6%. Phylogenetic results indicated that the pig has the closest relationship with cattle and dog. Two different nucleotides (A or G) at 95 nucleotide in the coding region of the PPARδ gene and three genotypes were found: AA, AB, BB. PPARδ mRNA expression in 14 tissues was tested by qPCR and the result showed that the expression was high in liver, spleen, ear and small intestine. This study provides experimental basis for further study on the expression and regulation of PPARδ gene. |
2楼2015-02-11 03:07:34