Affinity selection of cDNA libraries by λ phage surface display

被引:21
|
作者
Niwa, M
Maruyama, H
Fujimoto, T
Dohi, K
Maruyama, IN
机构
[1] Scripps Res Inst, Dept Cell Biol, La Jolla, CA 92037 USA
[2] Nara Med Univ, Dept Internal Med 1, Kashihara, Nara 6340813, Japan
关键词
autoantigen; bacteriophage vector; biopanning; fusion protein; Sjogren's syndrome;
D O I
10.1016/S0378-1119(00)00348-6
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Bacteriophage lambda surface display was used to isolate cDNA clones encoding autoantigens recognized by sera from patients with Sjogren's syndrome (SS). We made cDNA libraries from human HeLa and HepG2 cells, using the expression vector lambda foo. By repeating affinity selection of the libraries with the sera immobilized in microtiter wells, we isolated three clones that encode previously unknown antigens as well as four clones previously known as SS autoantigens. The newly identified autoantigens include TRK-fused gene product (TFG), survival motor neuron gene product (SMN) and pM5, which has a similarity to the metal-binding domain of human fibroblast collagenase. Thus, the bacteriophage lambda surface display is powerful for isolating cDNA clones by affinity screening. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:229 / 236
页数:8
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