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The use of recombinant DNA-based protein production using genetically modified plants could provide a reproducible, consistent quality, safe, animal-component free, origin-traceable, and cost-effective source for industrial proteins required in large amounts (1000s of metric tons) and at low cost (below US$100/Kg). The aim of this work was to demonstrate(More)
The polyprotein of Cocksfoot mottle virus (CfMV; genus SOBEMOVIRUS:) is translated from two overlapping open reading frames (ORFs) 2a and 2b by a -1 ribosomal frameshifting mechanism. In this study, a 12 kDa protein was purified from viral RNA-derived samples that appears to correspond to the CfMV genome-linked protein (VPg). According to the determined(More)
Processing of the polyprotein encoded by Potato virus A (PVA; genus Potyvirus) was studied using expression of the complete PVA polyprotein or its mutants from recombinant baculoviruses in insect cells. The time-course of polyprotein processing by the main viral proteinase (NIaPro) was examined with the pulse-chase method. The sites at the P3/6K1, CI-6K2(More)
The bioremediation potential of a nitrogen-fixing leguminous plant, Galega orientalis, and its microsymbiont Rhizobium galegae was evaluated in BTX (benzene, toluene, xylene)-contaminated soils in microcosm and mesocosm scale. To measure the intrinsic tolerance of the organisms to m-toluate, a model compound representing BTX, G. orientalis and R. galegae(More)
The polyprotein of Cocksfoot mottle virus (CfMV) is encoded by two overlapping open reading frames (ORFs). The putative replicase of CfMV is produced as a part of the polyprotein from ORF2b by the -1 ribosomal frameshifting mechanism. The signals leading to -1 ribosomal frameshifting directed by CfMV RNA are the slippery heptamer UUUAAAC and a stem-loop(More)
The ratio between proteins P27 and replicase of Cocksfoot mottle virus (CfMV) is regulated via a -1 programmed ribosomal frameshift (-1 PRF). A minimal frameshift signal with a slippery U UUA AAC heptamer and a downstream stem-loop structure was inserted into a dual reporter vector and directed -1 PRF with an efficiency of 14.4 +/- 1.9% in yeast and 2.4 +/-(More)
Introduction of sequences of interest into an intercistronic spacer (ICS) of dual reporter plasmids is the main experimental set-up used to identify and study internal ribosome entry sites (IRESs). We studied internal initiation of translation in yeast using the dicistronic approach. Three viral sequences and a polylinker-derived reference sequence were(More)
Comparison of the translational properties of Cocksfoot mottle virus 5'leader sequence with known translational enhancers from other plant viruses. (Manuscript) II Mäkeläinen K., and Mäkinen K. Testing of internal initiation via dicistronic constructs is complicated by production of extraneous transcripts. (Submitted) of VPg and the polyprotein processing(More)
Cover picture: Actin fi lament bundles stained with phalloidin in a sprouting bristle from a twinfi lin mutant pupa. This thesis is based on the following original publications, which are referred to in the text by their Roman numerals, and on unpublished data presented in the text.
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