학술논문

Expression of Agrobacterium rhizogenes rolBgene fusions in Escherichia coli: production of antibodies against the RolB protein
Document Type
Article
Source
Gene; March 1990, Vol. 87 Issue: 1 p139-143, 5p
Subject
Language
ISSN
03781119; 18790038
Abstract
Expression of the rolBgene of Agrobacterium rhizogenesTL-DNA is sufficient to trigger root differentiation in transformed plant cells. To investigate the role of RolB in differentiation, a large portion of rolB, comprising about 90% of its C-terminal coding sequence, was cloned into vectors pEX34 and pEA305 in frame with the truncated N termini of the pL-MS2 phage DNA polymerase and, respectively, the ptac-cIts phage λ repressor gene. Hybrid proteins were expressed from both fusions and the one from pMTBEX1 was utilized to raise antibodies. These antibodies specifically recognize the RolB moiety in both pL-MS2-rolBand ptac-cI-rolBfusions. Unfused, complete RolB protein was obtained by in vitro translation in a rabbit reticulocyte system of a transcript obtained by in vitro transcription of rolB. RolB protein is specifically immunoprecipitated by the antibodies raised against the hybrid protein MS2-RolB.