New highly specific anti-BnASY polyclonal antibody prepara-tion and application in immunofluorescence assay

摘要

ASY (asynaptic) is a synaptonemal complex (SC) related protein in plant mei-osis. Some ASYs in plants have been cloned and functional determined. ASY in Brassica napus has not been sequenced and functional certificated. Here, we first cloned ASY gene in Brassica napus (BnASY) and inserted it into vector pET-32a for expression fusion protein BnASY-6His in Escherichia coli BL21 (DE3). Purified fusion protein was used to produce polyclonal antibody. Specificity and application of polyclonal antibody was examined by Western blot and immunofluorescence assay. Results showed that BnASY-6His recombi-nant protein and anti-BnASY antibody were successfully obtained. Polyclonal anti-BnASY antibody had high specificity. Results of immunofluorescence showed that ASY signals appeared at leptotene stage, faded away gradually at pachytene stage and almost disap-peared at diplonema stage. In this study, highly specific polyclonal antibody was success-fully acquired and used for immunofluorescence assay in plant cells. It will contribute to functional research of ASY in B. napus.