pET Plasmids (plasmid Expression T7) are a family of vectors widely used for the high-level expression of recombinant proteins in Escherichia coli (E. coli). Developed by Studier and colleagues, the pET system utilizes the T7 RNA polymerase to drive the transcription of the gene of interest, allowing for tightly regulated and robust protein expression.

Content

  • pET Plasmids: Technical Insights
  • Structure and Components:
    • Origin of Replication (ori): pET plasmids contain the pBR322 origin of replication, ensuring a moderate copy number within the host cell, which helps in maintaining the stability of the plasmid.
    • Antibiotic Resistance Gene:
      • ampR or kanR: Provides resistance to ampicillin or kanamycin, allowing for the selection of bacteria that have taken up the plasmid.
    • T7 Promoter: A strong promoter recognized by T7 RNA polymerase, ensuring high-level transcription of the cloned gene.
    • Lac Operator (lacO): Positioned downstream of the T7 promoter, the lac operator allows for the regulation of gene expression by the lac repressor (LacI), which binds to the operator and prevents transcription in the absence of an inducer.
    • Multiple Cloning Site (MCS): A sequence containing several unique restriction enzyme sites, facilitating the insertion of the gene of interest.
    • RBS (Ribosome Binding Site): Located upstream of the cloning site, it ensures efficient translation initiation of the mRNA.
    • T7 Terminator: A sequence that signals the termination of transcription by T7 RNA polymerase, ensuring that the mRNA is properly processed.
  • Applications:
    • High-Level Protein Expression: pET plasmids are designed for robust expression of recombinant proteins in E. coli. Upon induction with IPTG, the T7 RNA polymerase drives high levels of transcription from the T7 promoter, leading to abundant protein production.
    • Tightly Regulated Expression: The presence of the lac operator allows for tight regulation of gene expression. In the absence of an inducer (IPTG), the lac repressor binds to the operator, preventing transcription. This minimizes leaky expression and potential toxicity of the recombinant protein.
    • Functional Studies: pET plasmids facilitate the production of large quantities of protein for biochemical, structural, and functional studies, including enzyme assays, crystallography, and interaction studies.
    • Industrial Applications: Due to their high expression levels, pET plasmids are used in the production of industrial enzymes, therapeutic proteins, and other biotechnological applications.
  • Types of pET Plasmids:
    • pET-3a, pET-5a, pET-9a, etc.: These early versions differ mainly in the arrangement of the MCS and the presence of different restriction sites.
    • pET-21, pET-28, pET-32, etc.: These vectors contain additional features such as N-terminal or C-terminal His-tags, allowing for easy purification of the expressed protein using metal affinity chromatography. Some also include other tags like S-tag or thioredoxin for enhanced solubility and detection.
    • pET-Duet, pET-Tricistronic, etc.: These specialized versions are designed for the co-expression of multiple genes, facilitating the study of protein-protein interactions and multi-subunit complexes.
  • Experimental Considerations:
    • Host Strain: The expression of proteins using pET plasmids requires a host strain that contains a chromosomal copy of the T7 RNA polymerase gene, such as BL21(DE3) or its derivatives.
    • Induction Conditions: Optimal induction conditions, including the concentration of IPTG, temperature, and duration, should be determined experimentally to maximize protein yield and solubility.
    • Purification: If the plasmid includes a His-tag or other affinity tag, purification can be achieved using metal affinity chromatography or other tag-specific methods. Verification of protein expression and purity can be performed using SDS-PAGE and Western blotting.
AffiPLASMID® pRSETB-ECFP-GABABR1a VTF-Ypet-2
CAT# AFG-NVF-111990
Size: 2 μg
198.10 198.1 USD
AffiPLASMID® pRSETB-ECFP-GABABR1a VTF-Ypet
CAT# AFG-NVF-111989
Size: 2 μg
466.00 466.0 USD
198.10 198.1 USD
276.00 276.0 USD
276.00 276.0 USD
AffiPLASMID® pET-SUMO-DFNA5-6XHis
CAT# AFG-NVF-096667
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-SUMO
CAT# AFG-NVF-096666
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-SacB Plasmid
CAT# AFG-NVF-096665
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-MthRnl
CAT# AFG-NVF-096664
Size: 2 μg
766.20 766.2 USD
AffiPLASMID® pET-MBP-NLS-Geo, st Plasmid
CAT# AFG-NVF-096663
Size: 2 μg
215.20 215.20000000000002 USD
AffiPLASMID® pETM30-HIP1R
CAT# AFG-NVF-096662
Size: 2 μg
163.90 163.9 USD
AffiPLASMID® pETM-30
CAT# AFG-NVF-096661
Size: 2 μg
215.20 215.20000000000002 USD
AffiPLASMID® pETM- 41
CAT# AFG-NVF-096660
Size: 2 μg
257.00 257.0 USD
AffiPLASMID® pETM- 11
CAT# AFG-NVF-096659
Size: 2 μg
215.20 215.20000000000002 USD
AffiPLASMID® pET-HSP65
CAT# AFG-NVF-096658
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pETG- 30A
CAT# AFG-NVF-096657
Size: 2 μg
257.00 257.0 USD
AffiPLASMID® pET-fasG
CAT# AFG-NVF-096656
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pETDuet-1-SopB-EGFP-Ssb-mCherry
CAT# AFG-NVF-096655
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pETDuet-1-SopB-EGFP
CAT# AFG-NVF-096654
Size: 2 μg
276.00 276.0 USD
276.00 276.0 USD
276.00 276.0 USD
276.00 276.0 USD
AffiPLASMID® pETDuet1-msyB-mcjA Plasmid
CAT# AFG-NVF-096650
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pETDuet-1 plasmid
CAT# AFG-NVF-096649
Size: 2 μg
182.90 182.9 USD
AffiPLASMID® pETDuet-1
CAT# AFG-NVF-096648
Size: 2 μg
182.90 182.9 USD
AffiPLASMID® pETDuet- 1 Plasmid
CAT# AFG-NVF-096647
Size: 2 μg
182.90 182.9 USD
AffiPLASMID® pET-dCas9-VP64-6xHis
CAT# AFG-NVF-096646
Size: 2 μg
198.10 198.1 USD
AffiPLASMID® pETconNK
CAT# AFG-NVF-096645
Size: 2 μg
466.00 466.0 USD
AffiPLASMID® pETCON-IgG1/design, 50
CAT# AFG-NVF-096644
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pETCON-HtsptLB12v.3
CAT# AFG-NVF-096643
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pETcon (-)
CAT# AFG-NVF-096642
Size: 2 μg
466.00 466.0 USD
AffiPLASMID® pETcoco- 1
CAT# AFG-NVF-096641
Size: 2 μg
214.25 214.25 USD
AffiPLASMID® pETBlue- 2
CAT# AFG-NVF-096640
Size: 2 μg
215.20 215.20000000000002 USD
AffiPLASMID® pET-6xHis- (-30) GFP
CAT# AFG-NVF-096637
Size: 2 μg
230.40 230.4 USD
AffiPLASMID® pET-6His-MBP-MpAgo
CAT# AFG-NVF-096636
Size: 2 μg
198.10 198.1 USD
AffiPLASMID® pET-5b (+)
CAT# AFG-NVF-096635
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-5a (+)
CAT# AFG-NVF-096634
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-50b (+)
CAT# AFG-NVF-096633
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-4CDS
CAT# AFG-NVF-096632
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-49b (+)
CAT# AFG-NVF-096631
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-48b (+)
CAT# AFG-NVF-096630
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-48b
CAT# AFG-NVF-096629
Size: 2 μg
230.40 230.4 USD
AffiPLASMID® pET-47b (+)
CAT# AFG-NVF-096628
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-46 EK/LIC
CAT# AFG-NVF-096627
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-45b (+) -Tn5
CAT# AFG-NVF-096626
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-45b (+)
CAT# AFG-NVF-096625
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-44c (+)
CAT# AFG-NVF-096624
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-44b (+)
CAT# AFG-NVF-096623
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-44a (+)
CAT# AFG-NVF-096622
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-44 EK/LIC
CAT# AFG-NVF-096621
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-43.1c (+)
CAT# AFG-NVF-096620
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-43.1b (+)
CAT# AFG-NVF-096619
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-43.1b Plasmid
CAT# AFG-NVF-096618
Size: 2 μg
215.20 215.20000000000002 USD
AffiPLASMID® pET-43.1a (+)
CAT# AFG-NVF-096617
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-43.1 EK/LIC
CAT# AFG-NVF-096616
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-42c (+)
CAT# AFG-NVF-096615
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-42b (+)
CAT# AFG-NVF-096614
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-42a (+)
CAT# AFG-NVF-096613
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-41Ek/LIC
CAT# AFG-NVF-096612
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-41c (+)
CAT# AFG-NVF-096611
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-41b (+)
CAT# AFG-NVF-096610
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET41a-IRFP720
CAT# AFG-NVF-096609
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-41a-DPEP1
CAT# AFG-NVF-096608
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET41a-CTNI
CAT# AFG-NVF-096607
Size: 2 μg
466.00 466.0 USD
AffiPLASMID® pET-41a (+)
CAT# AFG-NVF-096606
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-40b (+)
CAT# AFG-NVF-096605
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-3d
CAT# AFG-NVF-096604
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-3c-sumo
CAT# AFG-NVF-096603
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-3c Plasmid
CAT# AFG-NVF-096602
Size: 2 μg
257.00 257.0 USD
AffiPLASMID® pET3a-IL26 Plasmid
CAT# AFG-NVF-096601
Size: 2 μg
283.60 283.6 USD
AffiPLASMID® pET3a-FTH1-6×His
CAT# AFG-NVF-096600
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET39b-EK-Light
CAT# AFG-NVF-096599
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-39b (+)
CAT# AFG-NVF-096598
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-38b (+)
CAT# AFG-NVF-096597
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-37b (+)
CAT# AFG-NVF-096596
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-35b (+)
CAT# AFG-NVF-096595
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-33b (+)
CAT# AFG-NVF-096594
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-32c (+)
CAT# AFG-NVF-096593
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET-32b (+)
CAT# AFG-NVF-096592
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET32a-TP53
CAT# AFG-NVF-096591
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET32a-TEV
CAT# AFG-NVF-096590
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET32a-TatNLS-Runx2
CAT# AFG-NVF-096589
Size: 2 μg
276.00 276.0 USD
283.60 283.6 USD
AffiPLASMID® pET32a-Runx2-r
CAT# AFG-NVF-096587
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET32a-RbcS
CAT# AFG-NVF-096586
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET32a-RbcL
CAT# AFG-NVF-096585
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET32a-NR2C2 Plasmid
CAT# AFG-NVF-096584
Size: 2 μg
257.00 257.0 USD
AffiPLASMID® pET32a-MS2
CAT# AFG-NVF-096583
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET32a-mmADAMTS (254-553aa) Plasmid
CAT# AFG-NVF-096582
Size: 2 μg
283.60 283.6 USD
AffiPLASMID® pET32a-leptin Plasmid
CAT# AFG-NVF-096581
Size: 2 μg
283.60 283.6 USD
AffiPLASMID® pET32a-IRFP720
CAT# AFG-NVF-096580
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET32a-IL26
CAT# AFG-NVF-096579
Size: 2 μg
163.90 163.9 USD
AffiPLASMID® pET32a-IL25
CAT# AFG-NVF-096578
Size: 2 μg
163.90 163.9 USD
AffiPLASMID® pET32a-IL21
CAT# AFG-NVF-096577
Size: 2 μg
163.90 163.9 USD
AffiPLASMID® pET32a-GST-TMEM106B
CAT# AFG-NVF-096576
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET32a-Gene2-9×His Plasmid
CAT# AFG-NVF-096575
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET32a-EGFP-APP Plasmid
CAT# AFG-NVF-096574
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET32a-CUP1-2
CAT# AFG-NVF-096573
Size: 2 μg
276.00 276.0 USD
AffiPLASMID® pET32a-cap Plasmid
CAT# AFG-NVF-096572
Size: 2 μg
283.60 283.6 USD
AffiPLASMID® pET32a-APP-EGFP-6×His Plasmid
CAT# AFG-NVF-096571
Size: 2 μg
276.00 276.0 USD