Cloning of cell-specific secreted and surface proteins by subtractive antibody screening
Cloning of cell-specific secreted and surface proteins by subtractive antibody screening
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DOI：
10.1038/nbt0698-581
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发表时间：
1998-06-01
影响因子：
46.9
通讯作者：
Lodish, HF
中科院分区：
文献类型：
--
作者：
Scherer, PE;Bickel, PE;Lodish, HF
为了鉴定和克隆编码细胞或组织特异性分泌蛋白和表面蛋白的基因，针对来自靶细胞的表面或分泌蛋白的复杂混合物产生多克隆抗血清，然后免疫耗竭识别来自非靶细胞或组织的蛋白的抗体。耗尽的抗血清用于筛选噬菌体cDNA表达库。由于我们对脂肪细胞如何与其他细胞沟通感兴趣，我们使用这种方法克隆了编码在脂肪细胞分化过程中诱导的分泌蛋白和质膜蛋白的cDNA。我们描述了其中的几个，包括一种新的质膜相关蛋白，S3-12。
To identify and clone genes that encode cell-or tissue-specific secreted and surface proteins, a polyclonal antiserum was raised against a complex mixture of surface or secreted proteins from the target cell, followed by immunodepletion of antibodies that recognize proteins from a nontarget cell or tissue. The depleted antiserum is used to screen bacteriophage cDNA expression libraries. Because of our interest in how adipocytes communicate with other cells, we have used this method to clone cDNAs encoding secreted and plasma membrane proteins that are induced during adipocyte differentiation. We describe several of these, including a novel plasma membrane-associated protein, S3-12.