[Expression and immunization testing of fusion protein of Newcastle disease virus in leaf tissue of transgenic rice].

Abstract

Transgenic plant is an attractive bioreactor to produce recombinant protein, such as safe and economic vaccine. In present study, a plant expression binary vector pUNDV, containing the 1.7 kb fusion protein gene of Newcastle Disease Virus (NDV F) under the control of maize ubiquitin (Ubi) promoter and nos terminator was constructed, in which, the Hygromycin phosphotransferase (HPT) gene was used as the selectable marker. The expression cassette was introduced into a japonica rice variety by Agrobacterium-mediated transformation, and 6 independent transgenic rice lines were regenerated and verified by hygromycin resistance selection. The integration of target gene in transgenic plants was confirmed by PCR analysis. The results from ELISA and Western blot analyses revealed that NDV F could be expressed in the leaf of several transgenic plants. The total salt soluble proteins were extracted from leaf of transgenic plant F5, which contained the highest expressing level of NDV F protein, and used in immunization of BALB/c mice. The specific antibodies could be elicited in mice immunized with NDV F protein expressed in transgenic rice.