Difference between revisions of "Part:BBa K2230002"

 
 
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__NOTOC__
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<partinfo>BBa_K2230002 short</partinfo>
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[[File:Mingdaophil1025-5.png|300px]]
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===Erythromycin Resistance Gene as a Selection Marker for Lactobacillus transformation===
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Erythromycin is a common selection marker for engineering lactic acid bacteria. And teams [http://2013.igem.org/Team:Uppsala Uppsala] in 2013 and [http://2016.igem.org/Team:TMMU_China TMMU_China] in 2016 were using it to select the engineered Lactobacillus with erythromycin resistant strains. However, the BioBrick parts containing the erythromycin resistance gene are currently unavailable. On the other hand, iGEM HQ is unable to service the request of plasmid carrying erythromycin resistance gene. Maybe iGEM HQ can’t manipulate erythromycin resistant bacteria so far. Therefore, we decided to clone the erythromycin resistance gene (EmR) by ourselves. We searched the vector carrying EmR and found an iGEM team HKUST in 2010 working with it. We got the pMG36e vector from the team and cloned the EmR gene out onto pSB1C3 (EmR/pSB1C3, [https://parts.igem.org/Part:BBa_K2230001 BBa_K2230001]). We also made RBS-EmR/pSB1C3 ([https://parts.igem.org/Part:BBa_K2230002 BBa_K2230002]) and RBS-EmR-TT/pSB1C3 ([https://parts.igem.org/Part:BBa_K2230003 BBa_K2230003]) by assembling RBS (B0034) and double terminator (B0015) parts, respectively. These will provide iGEM team this resistance gene as selection marker in the future.
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===Cloning===
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EmR (Erythromycin resistance gene) was amplified from pMG36e vector and cut by XbaI & PstI. Then, the fragment was assembled with RBS/pSB1A2 (BBa_B0034) cut by SpeI & PstI
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===Reference===
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Genetic engineering techniques for lactic acid bacteria: construction of a stable shuttle vector and expression vector for β-glucuronidase. Biotechnol Lett. 2014;36(2):327-35.
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<!-- Add more about the biology of this part here
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2230002 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K2230002 parameters</partinfo>
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Latest revision as of 06:43, 25 October 2017


RBS-EmR/pSB1A2

Mingdaophil1025-5.png


Erythromycin Resistance Gene as a Selection Marker for Lactobacillus transformation

Erythromycin is a common selection marker for engineering lactic acid bacteria. And teams [http://2013.igem.org/Team:Uppsala Uppsala] in 2013 and [http://2016.igem.org/Team:TMMU_China TMMU_China] in 2016 were using it to select the engineered Lactobacillus with erythromycin resistant strains. However, the BioBrick parts containing the erythromycin resistance gene are currently unavailable. On the other hand, iGEM HQ is unable to service the request of plasmid carrying erythromycin resistance gene. Maybe iGEM HQ can’t manipulate erythromycin resistant bacteria so far. Therefore, we decided to clone the erythromycin resistance gene (EmR) by ourselves. We searched the vector carrying EmR and found an iGEM team HKUST in 2010 working with it. We got the pMG36e vector from the team and cloned the EmR gene out onto pSB1C3 (EmR/pSB1C3, BBa_K2230001). We also made RBS-EmR/pSB1C3 (BBa_K2230002) and RBS-EmR-TT/pSB1C3 (BBa_K2230003) by assembling RBS (B0034) and double terminator (B0015) parts, respectively. These will provide iGEM team this resistance gene as selection marker in the future.


Cloning

EmR (Erythromycin resistance gene) was amplified from pMG36e vector and cut by XbaI & PstI. Then, the fragment was assembled with RBS/pSB1A2 (BBa_B0034) cut by SpeI & PstI


Reference

Genetic engineering techniques for lactic acid bacteria: construction of a stable shuttle vector and expression vector for β-glucuronidase. Biotechnol Lett. 2014;36(2):327-35.


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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]