MOLECULAR-CLONING AND ANALYSIS OF FUNCTIONAL CDNA AND GENOMIC CLONES ENCODING BOVINE CELLULAR RETINOIC ACID-BINDING PROTEIN
MOLECULAR-CLONING AND ANALYSIS OF FUNCTIONAL CDNA AND GENOMIC CLONES ENCODING BOVINE CELLULAR RETINOIC ACID-BINDING PROTEIN
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DOI：
10.1073/pnas.84.16.5645
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发表时间：
1987-08-01
影响因子：
11.1
通讯作者：
MCCORMICK, AM
中科院分区：
文献类型：
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作者：
SHUBEITA, HE;SAMBROOK, JF;MCCORMICK, AM
从牛肾上腺cDNA文库中克隆了一个编码细胞视黄酸结合蛋白的重组cDNA克隆pCRABP-HS 1，转染COS-7细胞后，产生了一个具有生物学活性的视黄酸结合蛋白分子，分子量为15.5kDa。使用pCRABP-HS 1作为探针的RNA印迹杂交分析显示，在牛肾上腺、子宫和睾丸中，含有最高水平的视黄酸结合活性的组织中存在单个1050个核苷酸的mRNA种类。从卵巢、脾、肾或肝中提取的RNA中未检测到杂交，这些RNA含有相对低水平的细胞视黄酸结合蛋白活性。从EcoRI* 牛基因组文库中分离的基因组克隆的分析表明，牛细胞视黄酸结合蛋白基因由四个外显子和三个内含子组成。在克隆的该基因的5“序列中鉴定了两个推定的启动子序列。
A recombinant cDNA clone, pCRABP-HS1, encoding cellular retinoic acid-binding protein was isolated from a bovine adrenal cDNA library, COS-7 cells transfected with pCRABP-HS1 produced a biologically active retinoic acid-binding protein molecule of the expected molecular mass (15.5 kDa). RNA blot hybridization analysis using pCRABP-HS1 as a probe revealed a single 1050-nucleotide mRNA species in bovine adrenal, uterus, and testis, tissues that contain the highest levels of retinoic acid-binding activity. No hybridization was detected in RNA extracted from ovary, spleen, kidney, or liver, which contain relatively low levels of cellular retinoic acid-binding protein activity. Analysis of genomic clones isolated from an EcoRI* bovine genomic library demonstrated that the bovine cellular retinoic acid-binding protein gene is composed of four exons and three introns. Two putative promoter sequences were identified in the cloned 5'' sequence of the gene.