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编码大鼠肠道前载脂蛋白AIV信使核糖核酸一部分的互补脱氧核糖核酸的克隆

Cloning of a complementary deoxyribonucleic acid encoding a portion of rat intestinal preapolipoprotein AIV messenger ribonucleic acid.

作者信息

Gordon J I, Smith D P, Alpers D H, Strauss A W

出版信息

Biochemistry. 1982 Oct 26;21(22):5424-31. doi: 10.1021/bi00265a007.

Abstract

Apolipoprotein AIV is one of the principal apolipoproteins synthesized by the rat small intestine. We have cloned a cDNA encoding a portion of preapolipoprotein AIV mRNA. A kinetically fractionated cDNA probe highly enriched for the abundant intestinal mucosal mRNA sequences was used to screen a library of recombinants containing cDNA generated from total intestinal epithelial mRNA. The abundant class of mRNA sequences was defined by hybridization analyses. This frequency class had an aggregate complexity of 5300 nucleotides and represented 25% of accumulated mRNA sequences.l The mRNAs comprising this class were identified by in vitro translation and included preapolipoprotein AIV, preproapolipoprotein AI, intestinal fatty acid binding protein, and liver fatty acid binding protein. A cDNA-containing clone derived from preapolipoprotein AIV mRNA was identified among probe-positive recombinants. This cDNA was used to establish that apolipoprotein AIV mRNA has a mass of 550000 daltons (equivalent to 1780 nucleotides) and represents 0.013% of total cellular RNA in the fasting state. Acute feeding with triglyceride-rich meals resulted in a 2-fold increase in preapolipoprotein AiV mRNA after 4 h.

摘要

载脂蛋白AIV是大鼠小肠合成的主要载脂蛋白之一。我们克隆了一个编码前载脂蛋白AIV mRNA一部分的cDNA。一个通过动力学分级分离的、高度富集丰富的肠粘膜mRNA序列的cDNA探针,被用于筛选一个包含从总肠上皮mRNA产生的cDNA的重组体文库。通过杂交分析定义了丰富类别的mRNA序列。这个频率类别的总复杂度为5300个核苷酸,占累积mRNA序列的25%。通过体外翻译鉴定了构成该类别的mRNA,包括前载脂蛋白AIV、前原载脂蛋白AI、肠脂肪酸结合蛋白和肝脂肪酸结合蛋白。在探针阳性的重组体中鉴定出一个源自前载脂蛋白AIV mRNA的含cDNA克隆。该cDNA被用于确定载脂蛋白AIV mRNA的质量为550000道尔顿(相当于1780个核苷酸),在禁食状态下占总细胞RNA的0.013%。富含甘油三酯的餐食急性喂养4小时后,前载脂蛋白AIV mRNA增加了2倍。

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