Difference between revisions of "Part:BBa K777109"
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===Usage and Biology===
 
===Usage and Biology===
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[[Image:YhjH_motility.jpg|thumb|300px|right|'''Fig. 2:''' BL21 ''E. coli'' carrying different constructs on [http://2012.igem.org/Team:Goettingen/Project/Materials#Tryptone_swimming_agar tryptone swimming agar] after 12h incubation at 33°C. Cells expressing ''fliC'' in puc18 under the natural promoter travelled approximately 0.25cm (radius) whereas no swimming could be detected for the control plasmid carrying [https://parts.igem.org/Part:BBa_K777125 K777125]. On tryptone swimming agar ''yhjH'' transformants were usually faster.]]
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[[Image:YhjH_motility.jpg|thumb|500px|right|'''Fig. 2:''' BL21 ''E. coli'' carrying different constructs on [http://2012.igem.org/Team:Goettingen/Project/Materials#Tryptone_swimming_agar tryptone swimming agar] after 12h incubation at 33°C. Cells expressing ''fliC'' in puc18 under the natural promoter travelled approximately 0.25cm (radius) whereas no swimming could be detected for the control plasmid carrying [https://parts.igem.org/Part:BBa_K777125 K777125]. On tryptone swimming agar ''yhjH'' transformants were usually faster.]]
 
We expected that transformation of ''E. coli'' with ''fliC'' constructs would results in longer flagella filaments. Unfortunately we were not able to confirm this via transmission electron microscopy. However we tested the transformed strains in swimming agar assays and they were significantly faster than control strains (Fig. 2). This was the case on [http://2012.igem.org/Team:Goettingen/Project/Materials#M9_Swimming_Agar M9 minimal medium swarming agar] and also on [http://2012.igem.org/Team:Goettingen/Project/Materials#Tryptone_swimming_agar tryptone swimming agar]. This indicates that ''fliC'' transformation indeed caused longer flagella filaments and thus higher motility in BL21 ''E. coli''  
 
We expected that transformation of ''E. coli'' with ''fliC'' constructs would results in longer flagella filaments. Unfortunately we were not able to confirm this via transmission electron microscopy. However we tested the transformed strains in swimming agar assays and they were significantly faster than control strains (Fig. 2). This was the case on [http://2012.igem.org/Team:Goettingen/Project/Materials#M9_Swimming_Agar M9 minimal medium swarming agar] and also on [http://2012.igem.org/Team:Goettingen/Project/Materials#Tryptone_swimming_agar tryptone swimming agar]. This indicates that ''fliC'' transformation indeed caused longer flagella filaments and thus higher motility in BL21 ''E. coli''  
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 20:30, 26 September 2012

fliC

  • The fliC gene codes for flagellin. This protein is the subunit that forms the flagellar filament of E. coli.




Usage and Biology

Fig. 2: BL21 E. coli carrying different constructs on [http://2012.igem.org/Team:Goettingen/Project/Materials#Tryptone_swimming_agar tryptone swimming agar] after 12h incubation at 33°C. Cells expressing fliC in puc18 under the natural promoter travelled approximately 0.25cm (radius) whereas no swimming could be detected for the control plasmid carrying K777125. On tryptone swimming agar yhjH transformants were usually faster.

We expected that transformation of E. coli with fliC constructs would results in longer flagella filaments. Unfortunately we were not able to confirm this via transmission electron microscopy. However we tested the transformed strains in swimming agar assays and they were significantly faster than control strains (Fig. 2). This was the case on [http://2012.igem.org/Team:Goettingen/Project/Materials#M9_Swimming_Agar M9 minimal medium swarming agar] and also on [http://2012.igem.org/Team:Goettingen/Project/Materials#Tryptone_swimming_agar tryptone swimming agar]. This indicates that fliC transformation indeed caused longer flagella filaments and thus higher motility in BL21 E. coli






















Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1224
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 301
    Illegal AgeI site found at 709
  • 1000
    COMPATIBLE WITH RFC[1000]


Fig. 1: Plasmid map of K777109 in pSB1C3