Structure of the mouse myelin P2 protein gene.
Structure of the mouse myelin P2 protein gene.
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小鼠髓磷脂 P2 蛋白基因的结构。
DOI：
10.1111/j.1471-4159.1991.tb02101.x
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发表时间：
1991
影响因子：
4.7
通讯作者：
Tennekoon,GI
中科院分区：
文献类型：
--
作者：
Narayanan,V;Kaestner,KH;Tennekoon,GI
髓磷脂P2蛋白是一种小分子（14，800 Da）蛋白，存在于外周和中枢神经系统髓磷脂中。为了研究这种髓鞘蛋白的表达调控，用兔P2cDNA（pSN2.2-2）筛选小鼠基因组文库，获得了含有20 kb插入片段的单个阳性噬菌体克隆。该插入物包含一个内部Sailrestriction位点和几个EcoRI位点。将此插入片段的EcoRI片段亚克隆到Bluescript中。兔P2cDNA（pSN2.2-2）与4kb EcoRI片段杂交，然后在创建巢式缺失后对该4kb片段进行测序。小鼠基因含有四个外显子：外显子1编码氨基酸1-24，外显子2编码氨基酸24-81，外显子3编码氨基酸82-115，外显子4编码氨基酸116-131。3个内含子的长度分别为1.2kb、150 bp和1.3kb。引物延伸分析揭示了在+1和+47处的两个转录起始位点，与两个P2 mRNA的存在一致，其中较大的转录物显得更丰富。从小鼠DNA预测的氨基酸序列表明，小鼠蛋白质与兔蛋白质在16个不同的位置不同，其中大部分差异位于外显子3。当比较脂肪酸结合蛋白（FABP）基因的基因结构时，P2基因与FABP家族中的其他基因具有相同的整体结构，表明该家族成员具有共同的祖先基因。5′-侧翼区含有候选TATA和CAAT序列，以及两个AP-1结合位点，这可能对调节该基因的表达很重要。
Myelin P2protein is a small (14,800 Da) protein found in peripheral and central nervous system myelin. To investigate the regulation of expression of this myelin protein, a mouse genomic library was screened with a rabbit P2cDNA (pSN2.2–2), and a single positive phage clone containing a 20‐kb insert was obtained. This insert contained a single internalSailrestriction site and severalEcoRI sites. TheEcoRI fragments from this insert were subcloned into Bluescript. The rabbit P2cDNA (pSN2.2–2) hybridized with a 4‐kbEcoRI fragment, and this 4‐kb fragment was then sequenced after the creation of nested deletions. The mouse gene contained four exons: exon 1 coded for amino acids 1–24, exon 2 for amino acids 24–81, exon 3 for amino acids 82–115, and exon 4 for amino acids 116–131. The three introns were 1.2 kb, 150 bp, and 1.3 kb in length. Primer extension analysis revealed two transcription start sites at +1 and +47, consistent with the presence of two P2mRNAs, with the larger transcript appearing more abundant. The amino acid sequences predicted from the mouse DNA indicate that the mouse protein differs from the rabbit protein at 16 different positions, with most of the differences located in exon 3. When the gene structure of fatty acid binding protein (FABP) genes were compared, the P2gene had the same overall structure as others in the FABP family, suggesting a common ancestral gene for members of this family. The 5′‐flanking region contains candidate TATA and CAAT sequences, as well as two AP‐l binding sites that may be important in modulation of the expression of this gene.