Difference between revisions of "Part:BBa K2368021"

 
 
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<partinfo>BBa_K2368021 short</partinfo>
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<h1>Introduction</h1>
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<p style="text-align: center"><partinfo>BBa_K2368021 short</partinfo></p>
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The original G&#945; in the yeast is modified so that it can be adapted to human sweet receptors. We modified the base of the unmodified G&#945; at the carbon end.In addition to the transformation of the human base we also carried out other transformations on other species, and the carbon end of G&#945; was changed to the Schizosaccharomyces pombe sequence, and the The sweetness end of the receptor of G&#945;-carbon-terminated Schizosaccharomyces pombe sequence can be adapted thus the signal pathway can work properly.
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<p> Gα plays an important role in MAPK pathway. In the yeast endogenous metabolic pathway, the conformation of Ste2 changes and combines with the Gα, forcing Gα dissociates with Gβ/Gγ dimer. In order to make the sweetness receptor fit the Gα better, we modified the Gα. Then the sweetness receptor can combine with the Gα and send the signal to the downstream.</p>
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[[File:Ga-S-1-HR.png|center|500px|加载失败时候的说明文字]]
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<p style="text-align: center"> Fig.1 The structure of Gα in <i>Schizosaccharomyces pombe</i> </p>
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<h1>Design</h1>
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<p> Mutated C-terminus 15bp sequence of the yeast Gα, and replace it with the <i>Schizosaccharomyces pombe</i> Gα subunit. After construct the modified Gα by PCR, and the initial sequence is amplified from <i>Saccharomyces Cerevisiae CEN.PK2-1C</i>.</p>
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[[File: GS-1-HR.png|center|500px|加载失败时候的说明文字]]
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<p style="text-align: center"> Fig.2 The schematic diagram of Modified Gα-<i>Schizosaccharomyces pombe </i> </p>
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<h1>Experiment</h1>
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<p> The part is used to improve the matching condition of sweetness receptor and the modified Gα. The length of sequence is 1421bp.</p>
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[[File: 1GS-2-HR.png|center|500px|加载失败时候的说明文字]]
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<p style="text-align: center"> Fig.3 Electrophoresis of the positive result of the modified Gα- <i>Schizosaccharomyces pombe</i> </p>
 
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===Usage and Biology===
 
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<span class='h3bb'>Sequence and Features</span>
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<h2>Sequence and Features</h2>
 
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===Functional Parameters===
 
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<partinfo>BBa_K2368021 arameters</partinfo>
 
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Latest revision as of 11:07, 26 October 2017

Introduction

Modified Gα Schizosaccharomyces pombe

Gα plays an important role in MAPK pathway. In the yeast endogenous metabolic pathway, the conformation of Ste2 changes and combines with the Gα, forcing Gα dissociates with Gβ/Gγ dimer. In order to make the sweetness receptor fit the Gα better, we modified the Gα. Then the sweetness receptor can combine with the Gα and send the signal to the downstream.

加载失败时候的说明文字

Fig.1 The structure of Gα in Schizosaccharomyces pombe

Design

Mutated C-terminus 15bp sequence of the yeast Gα, and replace it with the Schizosaccharomyces pombe Gα subunit. After construct the modified Gα by PCR, and the initial sequence is amplified from Saccharomyces Cerevisiae CEN.PK2-1C.

加载失败时候的说明文字

Fig.2 The schematic diagram of Modified Gα-Schizosaccharomyces pombe

Experiment

The part is used to improve the matching condition of sweetness receptor and the modified Gα. The length of sequence is 1421bp.

加载失败时候的说明文字

Fig.3 Electrophoresis of the positive result of the modified Gα- Schizosaccharomyces pombe

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]