Tissue-specific expression of a cDNA clone from cell suspension cultures of Lupinus polyphyllus
Perrey, R.; Schneider, M.; Wink, M.
Plant Molecular Biology 14(6): 1055-1056
ISSN/ISBN: 0167-4412 PMID: 2102872 DOI: 10.1007/bf00019403
A cDNA library was constructed from poly(A)+ RNA from suspension-cultured cells derived from stem tissue. The cDNA synthesis was carried out using an RNase H approach and EcoRI adaptor strategy. The cDNA-adaptor construction was cloned in the EcoRI sites of the expression vector pUC19 and the cDNA library was established by transforming competent Escherichia coli DH5 alpha to produce 2.5 x 105 transformants/ micro g poly(A)+ RNA. The library was screened immunologically with polyclonal antibodies against horseradish peroxidase; 17 positive clones with inserts between 500 and 1500 bp were isolated and sequenced by the chain termination and/or chemical method.