Hi,
I have done yeast signal trap assay to identify secretary proteins. I have cloned my target gene (only one gene) into YST vector and transformed into invertase mutant yeast. When I culture the transformants on sucrose medium, about 50% transformants grow on sucrose medium and the rest do not grow. How to interpret this observation? Is the gene code a secretary protein? or something wrong with the transformation? Experts, please help me…
some developmental processes and induced plant responses have been identified that are directly or indirectly influenced by wall-localized, or apoplastic, molecular interactions and signalling pathways. The yeast-based signal sequence trap (YSST) is a potentially valuable experimental tool to characterize the proteome of the wall and apoplast, or 'secretome', although few studies have been performed with plants and to date this strategy has not been coupled with a subsequent analysis to confirm extracellular localization of candidate proteins in planta. This current report describes the use of the YSST, together with transient expression of a selection of identified proteins as fusions with the reporter GFP, focusing on the complex extracellular interactions between peach (Prunus persica) pollen and pistil tissues.
good luck
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