Analysis of the interaction between single-chain variable fragments and their antigen in a reducing intracellular environment using the two-hybrid system

被引：19

作者：

De Jaeger, G ^{[1
]}

Fiers, E ^{[1
]}

Eeckhout, D ^{[1
]}

Depicker, A ^{[1
]}

机构：

[1] State Univ Ghent VIB, Dept Plantengenet, Vakgrp Mol Genet, B-9000 Ghent, Belgium

来源：

FEBS LETTERS | 2000年 / 467卷 / 2-3期

关键词：

dihydroflavonol-4-reductase; two-hybrid; immunomodulation; reducing subcellular environment; single-chain variable fragment;

D O I：

10.1016/S0014-5793(00)01182-0

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The coding sequences of three single-chain variable (scFv) fragments (A4, G4 and H3), which bind to dihydroflavonol-4-reductase (DFR) of Petunia hybrida, and the DFR-encoding sequence were cloned in two-hybrid vectors. The vectors were transformed in the yeast strain HF7c (his3-200, trp1-901, leu2-3) and the scFv-DFR interaction was analyzed by measuring yeast growth on medium without histidine. ScFv-G4 and, to a lesser extent, scFv-A4 could interact with DFR in the east nucleus. On the contrary, scFv-H3 showed no interaction with its antigen in yeast. The results of a previous expression analysis of the same scFv Fragments in the plant cytosol correlate with those of the two-hybrid test. This suggests that it is possible to evaluate the antigen-scFv interaction in a reducing subcellular environment with the two-hybrid test. Therefore, the yeast two-hybrid system tan be useful to identify candidate scFv fragments for intracellular antibody applications. (C) 2000 Federation of European Biochemical Societies.

引用

页码：316 / 320

页数：5

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