Function analysis of 5'-UTR of the cellulosomal xyl -doc cluster in Clostridium papyrosolvens

Biotechnology for Biofuels
Xia ZouChenggang Xu

Abstract

Anaerobic, mesophilic, and cellulolytic Clostridium papyrosolvens produces an efficient cellulolytic extracellular complex named cellulosome that hydrolyzes plant cell wall polysaccharides into simple sugars. Its genome harbors two long cellulosomal clusters: cip-cel operon encoding major cellulosome components (including scaffolding) and xyl-doc gene cluster encoding hemicellulases. Compared with works on cip-cel operon, there are much fewer studies on xyl-doc mainly due to its rare location in cellulolytic clostridia. Sequence analysis of xyl-doc revealed that it harbors a 5' untranslated region (5'-UTR) which potentially plays a role in the regulation of downstream gene expression. Here, we analyzed the function of 5'-UTR of xyl-doc cluster in C. papyrosolvens in vivo via transformation technology developed in this study. In this study, we firstly developed an electrotransformation method for C. papyrosolvens DSM 2782 before the analysis of 5'-UTR of xyl-doc cluster. In the optimized condition, a field with an intensity of 7.5-9.0 kV/cm was applied to a cuvette (0.2 cm gap) containing a mixture of plasmid and late cell suspended in exponential phase to form a 5 ms pulse in a sucrose-containing buffer. Afterwards, the putativ...Continue Reading

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Datasets Mentioned

BETA
ACXX00000000.2
GSE57652

Methods Mentioned

BETA
restriction assay
PCR
restriction
electrophoresis
electrophoretic mobility shift assay
electron microscopy
restriction modification
Protein Assay
scanning electron microscopy

Software Mentioned

MEGA7
ViennaRNA package
RNAalifold

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