小鼠神经细胞黏附分子L1的cDNA克隆鉴定
Identification of cDNA clones of the mouse neural cell adhesion molecule L1.
作者信息
Tacke R, Moos M, Teplow D B, Früh K, Scherer H, Bach A, Schachner M
机构信息
Department of Neurobiology, University of Heidelberg, F.R.G.
出版信息
Neurosci Lett. 1987 Nov 10;82(1):89-94. doi: 10.1016/0304-3940(87)90176-5.
Two cDNA clones of the neural cell adhesion molecule L1 (Mr 200,000) were isolated using lambda gt10 and lambda gt11 libraries constructed from postnatal day 8 mouse brain poly(A)+ RNA. Clone K21 was selected and identified using immunoaffinity purified polyclonal antibodies. It was then used to isolate a secondary clone (K21-1), which hybridized with an oligonucleotide probe synthesized by reverse translation of the aminoterminal sequence of the 80 kDa carboxyterminal proteolytic fragment of L1. Blot hybridization analysis indicated that L1 is encoded by a single gene and transcribed by a single 6 kb mRNA which is present only in cells or tissues known to express L1.
利用从出生后第8天小鼠脑多聚腺苷酸加尾RNA构建的λgt10和λgt11文库，分离出神经细胞黏附分子L1（分子量200,000）的两个cDNA克隆。选择克隆K21并用免疫亲和纯化的多克隆抗体进行鉴定。然后用它分离出一个二级克隆（K21-1），该克隆与通过L1 80 kDa羧基末端蛋白水解片段氨基末端序列反向翻译合成的寡核苷酸探针杂交。印迹杂交分析表明，L1由单个基因编码，并由仅存在于已知表达L1的细胞或组织中的单个6 kb mRNA转录。
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