人肝癌cDNA表达文库中人类血清白蛋白和α-L-岩藻糖苷酶克隆的显色免疫检测
Chromogenic immunodetection of human serum albumin and alpha-L-fucosidase clones in a human hepatoma cDNA expression library.
作者信息
de Wet J R, Fukushima H, Dewji N N, Wilcox E, O'Brien J S, Helinski D R
出版信息
DNA. 1984 Dec;3(6):437-47. doi: 10.1089/dna.1.1984.3.437.
A human hepatoma cDNA library was constructed in lambda gt11, a bacteriophage vector that was designed to express cDNA-encoded antigenic determinants in Escherichia coli. The cDNA expression library contained approximately 8 X 10(6) recombinant phages with an average insert size of 780 bp. The feasibility of using a chromogenic immunodetection procedure to isolate cDNA clones was proved by isolating a human serum albumin (HSA) cDNA clone. An approximately 1.0-kb cDNA clone was then isolated by screening the library with rabbit anti-human alpha-L-fucosidase antibodies. The identity of the alpha-L-fucosidase cDNA clone was confirmed by DNA sequence analysis and a comparison of the predicted amino acid sequence to the amino acid sequences of human alpha-L-fucosidase tryptic peptides.
在λgt11噬菌体载体中构建了人肝癌cDNA文库，该噬菌体载体设计用于在大肠杆菌中表达cDNA编码的抗原决定簇。该cDNA表达文库包含约8×10⁶个重组噬菌体，平均插入片段大小为780 bp。通过分离人血清白蛋白（HSA）cDNA克隆，证明了使用显色免疫检测程序分离cDNA克隆的可行性。然后用兔抗人α-L-岩藻糖苷酶抗体筛选文库，分离出一个约1.0 kb的cDNA克隆。通过DNA序列分析以及将预测的氨基酸序列与人α-L-岩藻糖苷酶胰蛋白酶肽的氨基酸序列进行比较，证实了α-L-岩藻糖苷酶cDNA克隆的身份。
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