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인용 및 참조 문헌 (5)
Invitrogen™

pTrcHis2 TOPO™ TA Expression Kit

The pTrcHis- and pTrcHis2-TOPO™ TA Expression Kits allow fast, efficient cloning of PCR products into a prokaryotic expression vector for자세히 알아보기
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카탈로그 번호수량
K44000120 reactions
카탈로그 번호 K440001
제품 가격(KRW)
2,031,000
온라인 행사
Ends: 31-Mar-2026
2,389,000
할인액 358,000 (15%)
Each
수량:
20 reactions
제품 가격(KRW)
2,031,000
온라인 행사
Ends: 31-Mar-2026
2,389,000
할인액 358,000 (15%)
Each
The pTrcHis- and pTrcHis2-TOPO™ TA Expression Kits allow fast, efficient cloning of PCR products into a prokaryotic expression vector for high-level, regulated expression from the trc promoter. The vector in each kit, pTrcHis-TOPO™ or pTrcHis2-TOPO™, includes a minicistron element for enhanced translation efficiency of eukaryotic proteins in E. coli. Both vectors are provided linearized and activated with topoisomerase I for 5-minute TOPO™ Cloning with >85% cloning efficiency. To simplify protein purification and detection, the pTrcHis-TOPO™ and pTrcHis2-TOPO™ vectors include the following features:

pTrcHis-TOPO™

• N-terminal Xpress™ epitope for detection with an Anti-Xpress™ Antibody
• N-terminal polyhistidine (6xHis) tag for purification using nickelchelating resin and detection with an Anti-HisG Antibody
• Enterokinase cleavage site for efficient removal of N-terminal fusion tags

pTrcHis2-TOPO™
• C-terminal c-myc epitope for detection with an Anti-myc Antibody
• C-terminal polyhistidine (6xHis) tag for purification using nickel-chelating resin and detection with an Anti-His(C-term) Antibody
For Research Use Only. Not for use in diagnostic procedures.
사양
항생제 내성 박테리아Ampicillin (AmpR)
박테리아 또는 효모 균주TOP10
분열EK (Enterokinase) Recognition Site
구성 또는 유도성 시스템Inducible
발현 메커니즘Cell-Based Expression
발현 시스템E. coli
유도제IPTG
제품 유형TA Expression Kit
수량20 reactions
선택 제제(진핵)None
벡터pTrc
클로닝 방법TOPO-TA
제품라인TOPO
프로모터Trc, lacO
단백질 태그His Tag (6x), c-Myc Epitope Tag
Unit SizeEach
구성 및 보관
The pTrcHis- and pTrcHis2-TOPO™ TA Expression Kits each contain two boxes. The TOPO™ TA box contains 200 ng of linearized, topoisomerase I-activated pTrcHis-TOPO™ or pTrcHis2-TOPO™ vector, sterile water, dNTPs, 10X PCR buffer, a control template and primers, primers for sequencing or PCR screening, and an expression control plasmid. The One Shot™ TOP10 box contains twenty-one, 50-μl aliquots of chemically competent TOP10 E. coli, S.O.C. medium, and a supercoiled control plasmid. Store the TOPO TA Cloning™ box at -20°C. Store the One Shot™ box at -80°C. All reagents are guaranteed stable for 6 months when properly stored.

자주 묻는 질문(FAQ)

Can I store my competent E. coli in liquid nitrogen?

We do not recommend storing competent E. coli strains in liquid nitrogen as the extreme temperature can be harmful to the cells. Also, the plastic storage vials are not intended to withstand the extreme temperature and may crack or break.

How should I store my competent E. coli?

We recommend storing our competent E. coli strains at -80°C. Storage at warmer temperatures, even for a brief period of time, will significantly decrease transformation efficiency.

What are the advantages and/or disadvantages of using TOP10, DH5α, or other cloning strains versus BL21 Star (DE3) or BL21 (DE3) cells for expression with the pTrc system?

Top10, DH5α, other cloning strains

Advantages:
- Saves time, can go directly from cloning to expression.
- The glycerol stock is more stable because these strains are endA- and recA-.

Disdvantages:
- If GOI is toxic, the cloning step can be difficult.
- These cloning strains are not protease-deficient; therefore, the protein may be degraded.

BL21 Star(DE3) or BL21 (DE3)

Advantages:
- These expression strains are protease-deficient.
- You have to transform the plasmid into an expression strain.
- The glycerol stock may be unstable because these expression strains are not endA- and recA-.
- The (DE3) part is wasted because the promoter does not need T7 RNA polymerase.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

What competent cell strain can I use for expression with my pTrc Expression System?

The pTrc promoter is recognized by E. coli RNA polymerase, not T7 polymerase, and therefore can be expressed in any E. coli strain, not just BL21 strains. Therefore, you can use Top10, DH5α, etc. for expression. However, if your gene is toxic, the cloning step can be difficult as there is leakiness in expression.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

How does glucose repress the pTrc promoter?

A transcriptional activator protein called CAP (catabolite activator protein) normally binds upstream of the trc promoter and activates transcription. This protein requires cAMP to bind the DNA. Adding glucose to the medium can reduce intracellular cAMP levels. Supplementing LB medium and agar plates with glucose will repress basal level transcription from the trc promoter. We recommend that you include 25 mM, 0.5% glucose in the selection medium to ensure stability of your insert.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

View all pTrcHis2 TOPO™ TA Expression Kit FAQs

인용 및 참조 문헌 (5)

인용 및 참조 문헌
Abstract
Characterization of a Novel Drosophila melanogaster Galectin. EXPRESSION IN DEVELOPING IMMUNE, NEURAL, AND MUSCLE TISSUES.
Authors: Pace Karen E; Lebestky Tim; Hummel Thomas; Arnoux Pascal; Kwan Kent; Baum Linda G;
Journal:J Biol Chem
PubMed ID:11809773
'We have cloned and characterized the first galectin to be identified in Drosophila melanogaster. The amino acid sequence of Drosophila galectin showed striking sequence similarity to invertebrate and vertebrate galectins and contained amino acids that are crucial for binding beta-galactoside sugars. Confirming its identity as a galectin family member, the ... More
Kaposi's Sarcoma-Associated Herpesvirus/Human Herpesvirus 8 Transcriptional Activator Rta Is an Oligomeric DNA-Binding Protein That Interacts with Tandem Arrays of Phased A/T-Trinucleotide Motifs.
Authors:Liao W, Tang Y, Kuo YL, Liu BY, Xu CJ, Giam CZ,
Journal:J Virol
PubMed ID:12915555
'Kaposi''s sarcoma associated herpesvirus (KSHV)/human herpesvirus 8 (HHV-8) encodes an immediate early transcriptional activator, Rta, which mediates viral reactivation from latency and lytic viral replication. Here we report the purification and characterizations of HHV-8 Rta and its interaction with Rta-responsive DNA elements. The Rta response element (RtaRE) in the promoter ... More
Assimilation of nicotinamide mononucleotide requires periplasmic AphA phosphatase in Salmonella enterica.
Authors:Grose JH, Bergthorsson U, Xu Y, Sterneckert J, Khodaverdian B, Roth JR,
Journal:J Bacteriol
PubMed ID:15968063
'Salmonella enterica can obtain pyridine from exogenous nicotinamide mononucleotide (NMN) by three routes. In route 1, nicotinamide is removed from NMN in the periplasm and enters the cell as the free base. In route 2, described here, phosphate is removed from NMN in the periplasm by acid phosphatase (AphA), and ... More
Transition state analogue inhibitors of purine nucleoside phosphorylase from Plasmodium falciparum.
Authors: Kicska Gregory A; Tyler Peter C; Evans Gary B; Furneaux Richard H; Kim Kami; Schramm Vern L;
Journal:J Biol Chem
PubMed ID:11707439
'Immucillins are logically designed transition-state analogue inhibitors of mammalian purine nucleoside phosphorylase (PNP) that induce purine-less death of Plasmodium falciparum in cultured erythrocytes (Kicska, G. A., Tyler, P. C., Evans, G. B., Furneaux, R. H., Schramm, V. L., and Kim, K. (2002) J. Biol. Chem. 277, 3226-3231). PNP is present ... More
Characterization of monomeric L1 metallo-beta -lactamase and the role of the N-terminal extension in negative cooperativity and antibiotic hydrolysis.
Authors: Simm Alan M; Higgins Catherine S; Carenbauer Anne L; Crowder Michael W; Bateson John H; Bennett Peter M; Clarke Anthony R; Halford Stephen E; Walsh Timothy R;
Journal:J Biol Chem
PubMed ID:11940588
The L1 metallo-beta-lactamase from Stenotrophomonas maltophilia is unique among this class of enzymes because it is tetrameric. Previous work predicted that the two regions of important intersubunit interaction were the residue Met-140 and the N-terminal extensions of each subunit. The N-terminal extension was also implicated in beta-lactam binding. Mutation of ... More