Difference between revisions of "Part:BBa K2617006"

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<partinfo>BBa_K2617006 short</partinfo>
 
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Promoting folding of misfolded polypeptides and enhancing the tolerance of Escherichia coli to butanol.
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GroEL can promote folding of misfolded polypeptides and enhancing the tolerance of Escherichia coli to butanol. During the production process, the accumulation of butanol will exert a great inhibitory effect on cell growth. The GroEL gene and GroES gene from C. acetobutylicum to make escherichia coli heterologous expression of heat shock protein (a molecular chaperone), which has been shown to increase butanol production [1].In addition, It is a very good choice to use GroEL gene together with GroES gene [https://parts.igem.org/Part:BBa_K2617007 BBa_K2617007 ].
  
 
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==Characterization==
 
==Characterization==
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GroESL, which is a combination of GroEL [https://parts.igem.org/Part:BBa_K2617006 BBa_K2617006] and GroES [https://parts.igem.org/Part:BBa_K2617007 BBa_K2617007] gene has been shown to increase butanol production in E.coli [1].GroES and GroEL gene are combined with J23100 promoter. We have test the tolerance of E.coli BW25113 with GroESL gene and original BW25113 under the condition of 1% (v/V) butanol concentration. The result (Fig. 1) shows that GroESL has a positive effect on the butanol tolerance of E.coli.
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During the production process, the accumulation of butanol will exert a great inhibitory effect on cell growth. Therefore, we will introduce the GroEL gene and GroES gene from clostridium acetone butanol to make escherichia coli heterologous expression of heat shock protein -- a molecular chaperone, which has been shown to increase butanol production [1].To test butanol tolerance, the control strain DH5α, and the GroESL overexpression strain DH5α were grown in the presence of butanol in concentrations up to 1%.  
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[[File:T--UESTC-China--GroESL in BW25113.png|500px|thumb|center|'''Fig. 1''' After adding 0.3 ml butanol in 30 ml LB medium, the measurement of optical density (OD600) deference between BW25113 with GroESL and Negative Control in 24 hour.]]
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[[File:T--UESTC-China--GroESL in DH5α.png|500px|thumb|center|'''Fig. 1''' The measurement of optical density (A600) by periodically taking samples.]]
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===References===
 
===References===
  
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1.Zingaro, K. A., & Papoutsakis, E. T. (2013). GroESL overexpression imparts Escherichia coli tolerance to i-, n-, and 2-butanol, 1, 2, 4-butanetriol and ethanol with complex and unpredictable patterns. Metabolic engineering, 15, 196-205.
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[1]Abdelaal, A. S., Ageez, A. M., El, A. E. H. A. A., & Abdallah, N. A. (2015). Genetic improvement of n-butanol tolerance in Escherichia coli by heterologous overexpression of groESL operon from Clostridium acetobutylicum. 3 Biotech, 5(4), 401-410.
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K2617006 parameters</partinfo>
 
<partinfo>BBa_K2617006 parameters</partinfo>
 
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Latest revision as of 17:20, 30 September 2019


GroEL

GroEL can promote folding of misfolded polypeptides and enhancing the tolerance of Escherichia coli to butanol. During the production process, the accumulation of butanol will exert a great inhibitory effect on cell growth. The GroEL gene and GroES gene from C. acetobutylicum to make escherichia coli heterologous expression of heat shock protein (a molecular chaperone), which has been shown to increase butanol production [1].In addition, It is a very good choice to use GroEL gene together with GroES gene BBa_K2617007 .

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 880
    Illegal AgeI site found at 1341
  • 1000
    COMPATIBLE WITH RFC[1000]

Characterization

GroESL, which is a combination of GroEL BBa_K2617006 and GroES BBa_K2617007 gene has been shown to increase butanol production in E.coli [1].GroES and GroEL gene are combined with J23100 promoter. We have test the tolerance of E.coli BW25113 with GroESL gene and original BW25113 under the condition of 1% (v/V) butanol concentration. The result (Fig. 1) shows that GroESL has a positive effect on the butanol tolerance of E.coli.

Fig. 1 After adding 0.3 ml butanol in 30 ml LB medium, the measurement of optical density (OD600) deference between BW25113 with GroESL and Negative Control in 24 hour.

References

[1]Abdelaal, A. S., Ageez, A. M., El, A. E. H. A. A., & Abdallah, N. A. (2015). Genetic improvement of n-butanol tolerance in Escherichia coli by heterologous overexpression of groESL operon from Clostridium acetobutylicum. 3 Biotech, 5(4), 401-410.