Difference between revisions of "Part:BBa K3589151"

 
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<partinfo>BBa_K3589151 short</partinfo>
 
<partinfo>BBa_K3589151 short</partinfo>
  
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This basic part contains the sequence of an HA- as well as an 8xHis-tag (B5). Combined with a CDS, an appropriate promoter and a terminator your target protein should be detectable using an HA-antibody and you should be able to purify it using a nickel-NTA-column.
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This basic part contains the sequence of the HA-RGS-8His-tag (B5). Combined with a secretion signal of the Kaiser Collection from TU Kaiserslauterns 2019 iGEM Team like <a href="https://parts.igem.org/Part:BBa_K3002007">BBa_K3002007</a> (cCA (B2)), <a href="https://parts.igem.org/Part:BBa_K3002008">BBa_K3002008</a> (GLE (B2)) or <a href="https://parts.igem.org/Part:BBa_K3002009">BBa_3002009</a> (ARS (B2)), an appropriate promoter (<a href="https://parts.igem.org/Part:BBa_K3002001">BBa_K3002001 </a>(PSAD promoter (A1-B1)) or <a href="https://parts.igem.org/Part:BBa_K3002031">BBa_K3002031</a> (PAR promoter (A1-B1))) and a terminator (e.g. <a href="https://parts.igem.org/Part:BBa_K3002006">BBa_K3002006</a> (RPL23 terminator)) your target protein should be secreted and you should be able to detect it via HA-antibody and purify it via a nickel-NTA column.<br>
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Note that this part misses a SP20-portion. The SP20-portion is reported to improve the secretion of proteins by increasing the glycosylation (see Team TU Kaiserslautern 2019's <a href="https://2019.igem.org/Team:TU_Kaiserslautern/Results">results</a>). It is a repetition of Prolin and Serin, whereby Prolin is converted into hydroxyproline and then glycolyzed (<a href="https://doi.org/10.1111/pbi.12710">Ramos-Martinez <I>et. al</I>, 2017</a>).<br><br>
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<a href="https://parts.igem.org/Part:BBa_K3589150">BBa_K3589150</a> is a variation of this tag which possesses the SP20-portion.
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Revision as of 16:55, 25 October 2020


HA-RGS-8xHis tag for Chlamydomonas reinhardtii (Phytobrick)

This basic part contains the sequence of the HA-RGS-8His-tag (B5). Combined with a secretion signal of the Kaiser Collection from TU Kaiserslauterns 2019 iGEM Team like BBa_K3002007 (cCA (B2)), BBa_K3002008 (GLE (B2)) or BBa_3002009 (ARS (B2)), an appropriate promoter (BBa_K3002001 (PSAD promoter (A1-B1)) or BBa_K3002031 (PAR promoter (A1-B1))) and a terminator (e.g. BBa_K3002006 (RPL23 terminator)) your target protein should be secreted and you should be able to detect it via HA-antibody and purify it via a nickel-NTA column.
Note that this part misses a SP20-portion. The SP20-portion is reported to improve the secretion of proteins by increasing the glycosylation (see Team TU Kaiserslautern 2019's results). It is a repetition of Prolin and Serin, whereby Prolin is converted into hydroxyproline and then glycolyzed (Ramos-Martinez et. al, 2017).

BBa_K3589150 is a variation of this tag which possesses the SP20-portion.

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]