Plasmid backbones/Expression

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Plasmid backbones/Expression
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In molecular biology, researchers are often interested in expressing proteins for purification or characterization. Hence, several groups and companies have designed expression plasmids to make it easy to express recombinant proteins. Expression plasmids generally include a promoter and ribosome binding site for ready protein expression.

To express a protein coding sequence that adheres to the BioBrick assembly standard, simply assemble one of the parts below with your coding region of interest into any BioBrick plasmid backbone. Each of the parts below includes a promoter and a ribosome binding site for expression of a downstream protein coding region.

There are many plasmid backbones from which to choose. You might consider one of the high copy plasmid backbones for a plasmid backbone similar to pUC19. Or alternatively, you might consider a low or medium copy plasmid backbone, since lower copy number plasmids can in some cases give better protein expression.

BioBrickexpressionvector.png

Constitutive

If you're interested in constitutively expressing your protein of interest, use one of these plasmids.


There are no parts for this table


JeffTaborPhoto.jpg
Jeff Tabor constructed the composite expression part BBa_J13002 in Andy Ellington's lab. Note that if BBa_J13002 is combined with another part that expressed TetR repressor, then BBa_J13002 is inducible via anhydrous tetracycline or aTC.
SamanthaLiangPhoto.jpg
Samantha Liang, as a member of the 2006 UC Berkeley iGEM team, constructed the composite expression part BBa_J23038.

Inducible

If you're interested in controlled or inducible expression of your protein, use one of these plasmids. Generally, inducible expression plasmids only express your protein of interest upon addition of an exogenous inducer, such as IPTG, AHL, or arabinose.


More...
NameDescriptionResistanceRepliconCopy
number
ChassisLength
BBa_J04500IPTG inducible promoter with RBS    220
BBa_J130233OC6HSL+LuxR dependent POPS/RIPS generator    117
BBa_K081006Plambda promoter and RBS    72
BBa_K1065203Efe+Bba_B0015 in pSpac (BBa_K823026) A(E. coli) + K(B. subtilis)pDG148 B. subtilis + E. coli9512
BBa_K1065204Efe+Bba_B0015 in BBa_K823024 (pXyl) A(E. coli) S(B. subtilis)  B. subtilis + E. coli6017
BBa_K1362091pSB1A30: High copy BioBrick cloning/expression backbone carrying Amp resitanceApMB1100-300 2206
BBa_K1362095pSB4A50: Low copy BioBrick cloning/expression backbone carrying Amp resistanceApSC101~5 3446
BBa_K1362096pSB4C50: Low(?) copy BioBrick cloning/expression backbone carrying Cm resistanceCpSC101unsure 3408
BBa_K1362097Low copy BioBrick expression backbone carrying Kan resistanceKpSC101~5 3470
BBa_K1680026pETUE    3324
BBa_K215000R0011+B0034, strong IPTG-inducible promoter with strong RBS.    75
BBa_K2387032CpxR-eYFPn[1-154] and CpxR-eYPFc[155-238] + araC/pBAD promoterChloramphenicol  Escherichia coli3424
BBa_K2637009Gal2 promoter    535
BBa_K2637017CYC1 terminator    261
BBa_K2637059mutant Gal1 promoter    457
BBa_K2812002pBAD-ara promoter - Arabinose inducible regulatory promoter     71
BBa_K2812003Coding sequence for trunctated Lysostaphin regulated by T7-promoter    776
BBa_K2812004Coding sequence for trunctated Lysostaphin fused to His-tagged HlyA    1458
BBa_K2812005Coding sequence for trunctated Lysostaphin with HlyA and His6-tag regulated by T7-promoter    1496
BBa_K2812007Coding sequence for Pyocin S5 with HlyA and His6-tag regulated by pBAD-ara promoter    2288
BBa_K345667Lac regulated insertable promoter    220
BBa_K653003IPTG inducible Expression Platform    1083
BBa_K801004pTUM104 yeast shuttle vector with GAL1 promoter    5837
BBa_K823024pSBBs4S-Pxyl: Integrative expression vector for Bacillus subtilisA(E. coli) S(B. subtilis)  B. subtilis + E. coli4794
BBa_K823026pSBBs0K-Pspac (replicative Bacillus subtilis expression vector; IPTG inducibleA(E. coli) + K(B. subtilis)pDG148 B. subtilis + E. coli8289
BBa_S03511pLac : RBS    223


NoPhotoAvailable.jpg
Kristen DeCelle, as a member of the 2005 Davidson College iGEM team, constructed the IPTG-inducible composite expression part BBa_J04500. Note that without LacI-expression by the host chassis, BBa_J04500 is constitutive.
JeffTaborPhoto.jpg
Jeff Tabor constructed the AHL-inducible composite expression part BBa_J13023 in Andy Ellington's lab. Note that BBa_J13023 will only express your protein of interest if the activator LuxR is present and the repressor λ CI is absent.
2006DavidsonMissouriWesterniGEMteam.JPG
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The 2006 Davidson-Missouri Western joint iGEM team designed and constructed the composite expression part BBa_S03511. Note that without LacI-expression by the host chassis, BBa_S03511 is constitutive.

References

<biblio>

  1. Bernard01 pmid=18429183

</biblio>