【摘要】：試驗旨在克隆獲得PDK4、FGF10基因,并研究大白豬與從江香豬不同組織中PDK4、FGF10基因mRNA的表達差異。采用RT-PCR分別克隆從江香豬PDK4、FGF10基因并進行生物信息學(xué)分析,利用實時熒光定量PCR技術(shù)檢測PDK4、FGF10基因在大白豬和從江香豬不同組織中mRNA的相對表達量。結(jié)果顯示,從江香豬PDK4基因的編碼區(qū)全長1 224bp,編碼407個氨基酸;FGF10基因的編碼區(qū)全長636bp,編碼211個氨基酸。經(jīng)BLAST軟件進行同源性比對,發(fā)現(xiàn)從江香豬PDK4基因與羊、馬、人的核苷酸序列同源性分別為93%、92%和91%;FGF10基因與羊、牛、人、鼠的核苷酸序列同源性分別為94%、93%、93%和90%。由PDK4基因系統(tǒng)進化樹可知,從江香豬與牛、綿羊親緣關(guān)系較近;由FGF10基因系統(tǒng)進化樹可知,從江香豬與綿羊、牛、人、獼猴親緣關(guān)系較近,與小鼠和雞親緣關(guān)系較遠(yuǎn)。實時熒光定量PCR結(jié)果顯示,在從江香豬不同組織中,PDK4基因在腎臟中的表達量最高,在胃和脂肪中表達量較高,FGF10基因在胃中表達量最高,在腎臟和脂肪中表達量較高,兩個基因在背最長肌中的表達量均最低;在大白豬的不同組織中,PDK4、FGF10基因在脂肪中的表達量均最高,PDK4基因在背最長肌中的表達量最低,而FGF10基因在心臟中表達量最低。本試驗成功克隆了從江香豬PDK4、FGF10基因,并檢測了其在大白豬與從江香豬不同組織中的表達,為進一步研究PDK4、FGF10基因在脂質(zhì)代謝及脂肪沉積等方面的調(diào)控作用提供科學(xué)依據(jù)。
[Abstract]:The aim of the experiment was to clone and obtain PDK4,FGF10 gene and to study the difference of mRNA expression of PDK4,FGF10 gene in different tissues of big white pig and Congjiang Xiang pig. The PDK4,FGF10 gene of Congjiang Xiang pig was cloned by RT-PCR and analyzed by bioinformatics. The relative expression of PDK4,FGF10 gene in different tissues of big white pig and Congjiang Xiang pig was detected by real-time fluorescence quantitative PCR technique. The results showed that the coding region of PDK4 gene of Congjiang Xiang pig was 1224bpand the encoding region of 407 amino acid FGF10 gene was 636bp, encoding 211 amino acids. The nucleotide sequence homology of PDK4 gene of Congjiang Xiang pig and sheep, horse and human were 932% and 91% respectively. The homology of nucleotide sequence between PDK4 gene and sheep, cattle, human and mouse were 94 933% and 90%, respectively. According to the phylogenetic tree of PDK4 gene, Congjiang Xiang pig was closely related to cattle and sheep, and FGF10 gene phylogenetic tree showed that Congjiang Xiang pig was closely related to sheep, cattle, human and macaque, but far from mice and chicken. The results of real-time fluorescence quantitative PCR showed that the expression of PDK4 gene was the highest in kidney, the highest in stomach and fat, and the highest in kidney and fat. The expression of two genes in longissimus dorsi muscle was the lowest, and the expression of PDK4FGF10 gene in fat was the highest in different tissues of big white pig. The expression of PDK4 gene in longissimus dorsi muscle was the lowest, while that of FGF10 gene was the lowest in heart. In this experiment, the PDK4,FGF10 gene of Congjiang Xiang pig was cloned and its expression in different tissues of big white pig and Congjiang Xiang pig was detected, which provides scientific basis for further study on the regulation of PDK4,FGF10 gene in lipid metabolism and fat deposition.
【作者單位】： 貴州大學(xué)高原山地動物遺傳育種與繁殖省部共建教育部重點實驗室;貴州省動物遺傳育種與繁殖重點實驗室;貴州大學(xué)動物科學(xué)學(xué)院;貴州大學(xué)生命科學(xué)學(xué)院;
【基金】：國家科技支撐計劃(2015BAD03B02-3) 黔科合重大專項(黔科合NY字[2013]6008號)
【分類號】：S828

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1 鞠妍;劉華淼;魏海軍;;梅花鹿FGF10基因的生物信息學(xué)分析[J];遼寧師范大學(xué)學(xué)報(自然科學(xué)版);2014年01期

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本文編號：2203747