Difference between revisions of "Part:BBa K1951001:Design"
 
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__NOTOC__
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===Sequence optimization===
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<partinfo>BBa_K1951001 short</partinfo>
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<partinfo>BBa_K1951001 SequenceAndFeatures</partinfo>
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Snapgene has been used to remove the forbidden sites by changing in the substitution which didn't change the amino acid. The sequence has been sythetised by IDT<ref>http://eu.idtdna.com/site</ref>.
  
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===Codon optimization for <i>Escherichia coli</i>. ===
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We obtimised codon for <i>Escherichia coli</i>. Codon optimization is a technical used to <u>improve the protein expression</u> in living organism by increasing the translational efficiency of gene of interest [1-4, 6-13, 15-18, 20-28]. This biobrick codon optimised <u>increase the functionality of gene</u>. To process it, we use codon optimization IDT software<ref> https://eu.idtdna.com/CodonOpt </ref>.
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If you use this biobrick in <i>Escherichia coli</i>, you can be sure that the protein produced will be highly expressed and well solubilised.
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=== Prefix and suffix addition===
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Prefix and suffix subsequences (containing restriction site EcoRI, XbaI and SpeI PstI respectively) have been added by a SLIC method with the following oligos :
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{| class="wikitable"
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| desB slic forward 
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| cgctaaggatgatttctgGAATTCGCGGCCGCTTCTAGATGGGTATTGGTCTTGGG
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|-
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| desA slic reverse
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| ttgcccttttttgccggaCTGCAGCGGCCGCTACTAGTATTATTACACTGCAAATTC
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|}.
  
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===Design Notes===
 
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We had prefix and suffix to the DNA sequence containing restriction sites (EcoRI, Xba1 and SpeI, PstI respectively.
 
  
 
===Source===
 
===Source===

Latest revision as of 13:33, 17 October 2016

Sequence optimization

Snapgene has been used to remove the forbidden sites by changing in the substitution which didn't change the amino acid. The sequence has been sythetised by IDT[1].

Codon optimization for Escherichia coli.

We obtimised codon for Escherichia coli. Codon optimization is a technical used to improve the protein expression in living organism by increasing the translational efficiency of gene of interest [1-4, 6-13, 15-18, 20-28]. This biobrick codon optimised increase the functionality of gene. To process it, we use codon optimization IDT software[2]. If you use this biobrick in Escherichia coli, you can be sure that the protein produced will be highly expressed and well solubilised.

Prefix and suffix addition

Prefix and suffix subsequences (containing restriction site EcoRI, XbaI and SpeI PstI respectively) have been added by a SLIC method with the following oligos :

desB slic forward cgctaaggatgatttctgGAATTCGCGGCCGCTTCTAGATGGGTATTGGTCTTGGG
desA slic reverse ttgcccttttttgccggaCTGCAGCGGCCGCTACTAGTATTATTACACTGCAAATTC
.


Source

The following website contains the gene reference and its chromosomal location. http://strepdb.streptomyces.org.uk/cgi-bin/dc3.pl?accession=AL645882&serial=2769&width=900&start=3031362&end=3041362&iorm=map Start : 3037106 End : 3038347 The coding sequence of DNA comes from http://www.ncbi.nlm.nih.gov/nuccore/NC_003888.3

References

  1. ↑ http://eu.idtdna.com/site
  2. ↑ https://eu.idtdna.com/CodonOpt