Team:Michigan/Project
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== Oil Sands: Ag43 Expression == | == Oil Sands: Ag43 Expression == | ||
- | Since the NA degradation pathway in ''Pseudomonas'' is unknown, we decided to improve the biofilm formation abilities of ''Pseudomonas'' based on an early biofilm formation [[Team:Michigan/Oil_Sands_August_September#8/8/2010|crystal violet (CV) assay]]. We decided to clone the Antigen 43 (Ag43) self-dimerizing membrane complex from ''E. coli'', which has been shown to increase the adhesiveness of ''P. fluorescens'' and cause it to form co-species biofilms with ''E. coli'' [8]. The phenotype of ''P. fluorescens'' expressing Ag43 is a "frizzy" colony morphology, increased cell aggregation, and increased adhesiveness to glass surfaces. We hypothesized that this phenotype would improve the ability to use the ''Pseudomonas'' strains (LD1 and LD2) in an immobilized cell reactor (ICR), which has been shown to increase NA degradation 100 fold [9]. | + | Since the NA degradation pathway in ''Pseudomonas'' is unknown, we decided to improve the biofilm formation abilities of ''Pseudomonas'' based on an early biofilm formation [[Team:Michigan/Oil_Sands_August_September#8/8/2010|crystal violet (CV) assay]]. We decided to clone the ''flu'' operon, which expresses Antigen 43 (Ag43)- a self-dimerizing membrane complex from ''E. coli'', which has been shown to increase the adhesiveness of ''P. fluorescens'' and cause it to form co-species biofilms with ''E. coli'' [8]. The phenotype of ''P. fluorescens'' expressing Ag43 is a "frizzy" colony morphology, increased cell aggregation, and increased adhesiveness to glass surfaces. We hypothesized that this phenotype would improve the ability to use the ''Pseudomonas'' strains (LD1 and LD2) in an immobilized cell reactor (ICR), which has been shown to increase NA degradation 100 fold [9]. |
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magnification. [8]|500px]] | magnification. [8]|500px]] | ||
- | [[Image:Ag43 Flocs.png|left|FIG. 8. Settling of cells from | + | [[Image:Ag43 Flocs.png|left|thumb|FIG. 8. Settling of cells from |
static liquid suspensions. (A) E. coli HEHA16(pKKJ101), (B) E. coli | static liquid suspensions. (A) E. coli HEHA16(pKKJ101), (B) E. coli | ||
HEHA16(pUCP22), (C) P. fluorescens SBW25(pKKJ101), (D) P. fluorescens | HEHA16(pUCP22), (C) P. fluorescens SBW25(pKKJ101), (D) P. fluorescens | ||
SBW25(pUCP22), (E) K. pneumoniae(pKKJ101), and (F) K. pneumoniae( | SBW25(pUCP22), (E) K. pneumoniae(pKKJ101), and (F) K. pneumoniae( | ||
- | pUCP22). [8]|500px]] | + | pUCP22). Note plasmid pKKJ101 expresses the ''flu'' operon. [8]|500px]] |
<br style="clear: both" /> | <br style="clear: both" /> |
Revision as of 02:43, 26 October 2010