A scFv-alkaline phosphatase fusion protein which detects potato leafroll luteovirus in plant extracts by ELISA

被引：36

作者：

Harper, K ^{[1
]}

Kerschbaumer, RJ ^{[1
]}

Ziegler, A ^{[1
]}

Macintosh, SM ^{[1
]}

Cowan, GH ^{[1
]}

Himmler, G ^{[1
]}

Mayo, MA ^{[1
]}

Torrance, L ^{[1
]}

机构：

[1] UNIV AGR VIENNA,INST APPL MICROBIOL,VIENNA,AUSTRIA

来源：

JOURNAL OF VIROLOGICAL METHODS | 1997年 / 63卷 / 1-2期

关键词：

alkaline phosphatase fusion proteins; ELISA; phage display antibody library; potato leafroll luteovirus; scFv; virus detection;

D O I：

10.1016/S0166-0934(96)02133-7

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A single chain Fv antibody fragment (scFv) was obtained from a synthetic phage-antibody library after four rounds of selection against purified preparations of potato leafroll luteovirus (PLRV). Nucleotide sequence analysis showed that the scFv belongs to the human V(H)3 family. DNA encoding the scFv was sub-cloned into pDAP2 such that a scFv-alkaline phosphatase fusion protein was produced by transformed bacteria Following induction by isopropyl-beta-D-thiogalactopyranoside (IPTG). The fusion protein was obtained at concentrations of 10 mg/l of Escherichia coli culture medium and these fusion protein preparations were used directly in ELISA to detect PLRV in sap extracts from infected plants. Our work is the first report of the selection of a scFv specific for a luteovirus from a synthetic phage-display library and the production of a fusion protein with alkaline phosphatase for the detection of PLRV in infected plants. The results demonstrate the potential of scFv and enzyme-scFv fusion proteins in routine testing for plant virus infection. Copyright (C) 1997 Elsevier Science B.V.

引用

页码：237 / 242

页数：6

共 12 条

[1] ENHANCEMENT OF RESISTANCE TO POTATO LEAFROLL VIRUS MULTIPLICATION IN POTATO BY COMBINING THE EFFECTS OF HOST GENES AND TRANSGENES [J].