Isolation and characterisation of three moss-derived beta-tubulin promoters suitable for recombinant expression.
The moss Physcomitrella patens is an excellent tool to study plant gene-function relationships due to its high rate of homologous recombination (HR). It has also been shown to be very useful in the production of recombinant proteins which are secreted into a simple medium. Thus, there is a need for suitable promoters functional in this well established model organism. We isolated genomic flanking regions of the beta-tubulin gene family from Physcomitrella, concentrating on those family members showing high transcript abundance integrated over gametophytic tissues. Using a novel, fast and reliable quantification assay based on the transient expression and secretion of a recombinant human protein, three genomic upstream regions were characterised in serial deletion constructs. Expression rates were up to three times higher than those obtained with the 35S cauliflower mosaic virus (35S) promoter, which served as a reference.
Jost
Wolfgang
W
Greenovation Biotech GmbH, Bötzinger Strasse 29b, 79111 Freiburg, Germany.
Link
Sandra
S
Horstmann
Verena
V
Decker
Eva L
EL
Reski
Ralf
R
Gorr
Gilbert
G
D020121:5' Untranslated Regions; D001483:Base Sequence; D044002:Bryophyta / Q000235:genetics*; D003001:Cloning, Molecular; D020224:Expressed Sequence Tags; D017930:Genes, Reporter; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D011995:Recombination, Genetic; D012689:Sequence Homology, Nucleic Acid; D014404:Tubulin / Q000235:genetics*
D020121:5' Untranslated Regions; D012333:RNA, Messenger; D014404:Tubulin