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PPARD基因在動(dòng)物脂肪代謝過程中起重要作用，本研究以豬為實(shí)驗(yàn)動(dòng)物，通過RT-PCR方法擴(kuò)增得到PPARD基因全長(zhǎng)編碼區(qū)序列，在編碼區(qū)第95位檢測(cè)到G/A兩種核苷酸，序列提交GenBank數(shù)據(jù)庫獲得登錄號(hào)“KP757025和KP757026”。生物信息學(xué)分析顯示：該基因編碼462個(gè)氨基酸，蛋白分子量為49.74 kD，等電點(diǎn)為7.53。蛋白質(zhì)結(jié)構(gòu)分析PPARD存在2個(gè)保守域，無跨膜結(jié)構(gòu)，不存在信號(hào)肽，存在1個(gè)亮氨酸富集的核輸出信號(hào)，其N端、C端均親水；亞細(xì)胞定位顯示該蛋白分泌到細(xì)胞周質(zhì)的概率為69.6%。系統(tǒng)進(jìn)化分析表明，豬與牛、家犬的親緣關(guān)系較近。利用qPCR檢測(cè)了PPARD基因mRNA在豬14個(gè)組織中的表達(dá)量，發(fā)現(xiàn)在肝臟、耳朵、脾臟、小腸中表達(dá)量較高。本研究為進(jìn)一步研究豬PPARD基因的表達(dá)調(diào)控奠定研究基礎(chǔ)。
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: 金幣+100, 翻譯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.

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2樓2015-02-11 03:07:34