Difference between revisions of "Part:BBa K4339001"

 
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Alanine tRNA synthetase is an enzyme that catalyses the binding of alanine to tRNA. Modelling demonstrated that the expression of spider silk proteins (spidroins), which have a number of repetitive alanine regions, would be increased with the co-expression of alanine tRNA synthetase.
 
Alanine tRNA synthetase is an enzyme that catalyses the binding of alanine to tRNA. Modelling demonstrated that the expression of spider silk proteins (spidroins), which have a number of repetitive alanine regions, would be increased with the co-expression of alanine tRNA synthetase.
  
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===Usage and Biology===
 
===Usage and Biology===
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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The sequence originates from <i>E. coli</i> MG1655 K-12 (Puteny <i>et al.</i> 1981) 
 
<partinfo>BBa_K4339001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4339001 SequenceAndFeatures</partinfo>
  
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The enzyme was intended for co-expression with MaSps (Major Ampullate Spidroin Proteins) as part of a helper plasmid, containing genes for alanine tRNA and CycA (<partinfo>BBa_K4339000</partinfo> - an alanine import channel protein) expression. This system was designed to increase alanine uptake and synthesis of alanyl-tRNA to maximise possible rate of alanine incorporation into protein and thus, production of alanine-rich MaSps.
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This part was successfully synthesised but was not transformed due to time constraints.
  
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===References===
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Putney S et al. Purification and properties of alanine tRNA synthetase from <i>Escherichia coli</i>. A tetramer of identical subunits. <i>Journal of Biological Chemistry</i>. 1981;256(1): 198–204. doi: https://doi.org/10.1016/s0021-9258(19)70119-7
 
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K4339001 parameters</partinfo>
 
<partinfo>BBa_K4339001 parameters</partinfo>
 
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Latest revision as of 08:55, 12 October 2022


Alanine tRNA synthetase

Alanine tRNA synthetase is an enzyme that catalyses the binding of alanine to tRNA. Modelling demonstrated that the expression of spider silk proteins (spidroins), which have a number of repetitive alanine regions, would be increased with the co-expression of alanine tRNA synthetase.


Usage and Biology

Sequence and Features

The sequence originates from E. coli MG1655 K-12 (Puteny et al. 1981)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 2066
    Illegal PstI site found at 2582
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2066
    Illegal PstI site found at 2582
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 629
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 2066
    Illegal PstI site found at 2582
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 2066
    Illegal PstI site found at 2582
    Illegal AgeI site found at 709
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1090
    Illegal SapI.rc site found at 1138
    Illegal SapI.rc site found at 1288

The enzyme was intended for co-expression with MaSps (Major Ampullate Spidroin Proteins) as part of a helper plasmid, containing genes for alanine tRNA and CycA (BBa_K4339000 - an alanine import channel protein) expression. This system was designed to increase alanine uptake and synthesis of alanyl-tRNA to maximise possible rate of alanine incorporation into protein and thus, production of alanine-rich MaSps.

This part was successfully synthesised but was not transformed due to time constraints.

References

Putney S et al. Purification and properties of alanine tRNA synthetase from Escherichia coli. A tetramer of identical subunits. Journal of Biological Chemistry. 1981;256(1): 198–204. doi: https://doi.org/10.1016/s0021-9258(19)70119-7