Product Class: Other

pTXB1 Vector

Product Introduction

pTXB1 is an E. coli expression vector designed for the in-frame insertion of a target gene into the polylinker upstream of the Mxe intein/chitin binding domain. The fusion protein is bound to chitin beads and the thiol-induced cleavage activity of the intein releases the target protein.

  • View sequence details
  • Used in the IMPACT™ Protein Purification System
  • Recommended for use in intein-mediated protein ligation and C-terminal labeling
  • Double-stranded vector; 6,706 base pairs in length
  • Ampicillin resistance

Catalog # Size Concentration
N6707S 10.0 µg 200 µg/ml

Product Information

Description

IMPACT Citations

pTXB1 is an E. coli expression vector in the IMPACT™ Kit (NEB #E6901) (1,2). It is designed for the in-frame insertion of a target gene into the polylinker upstream of the Mxe intein/chitin binding domain (27 kDa) (2,3,4,5). The fusion protein is bound to chitin beads and the thiol-induced cleavage activity of the intein releases the target protein. pTXB vectors are recommended for use in intein-mediated protein ligation and C-terminal labeling (2). This double stranded vector is 6,706 base pairs in length.

DNASU and Addgene are central repositories for plasmid clones and collections that may also be helpful.

This product is related to the following categories:
IMPACT System,
Bacterial E. coli Protein Expression Products,
Chitin Purification (CBD-tag),
DNA Plasmids & Substrates Products,
Protein Purification Products,
Protein Expression Products,
This product can be used in the following applications:
Protein Expression in E. Coli,
Protein Expression

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Lac Repressor on Plasmid

0

Affinity Tag

Chitin-Binding Domain (CBD)

Features

  • The NdeI site in the polylinker contains an ATG sequence for translation initiation. Unique sites are indicated in bold. SalI site is not unique.
  • The SapI site should be used for cloning of the 3´ end of the insert. Use of the SapI site allows cloning of the target protein adjacent to the intein, resulting in cleavage of the target protein without any additional amino acids at its C-terminus.
  • Expression of the fusion gene is under the control of an IPTG-inducible T7 promoter (6).
  • A pBR322 derivative with a ColE1replication origin.
  • Origin of DNA replication from bacteriophage M13, which allows for the production of single-stranded DNA by helper phage superinfection of cells bearing the plasmid (M13K07 helper phage, NEB #N0315).
  • Ampicillin resistance.
  • Other IMPACT vectors are available which allow for fusion of a target gene to N- or C- terminus of an intein. The cleavage reaction may be induced by thiol reagent or temperature/pH shift.
  • Companion vector pTXB3 (NEB #N6708) contains an NcoI site in place of NdeI.
  • A wide range of E. coli host strains: T7 Express Competent E. coli (High Efficiency) (NEB #C2566) or BL21(DE3) Competent E. coli (NEB #C2527) and derivatives.

References

  1. Chong, S., Mersha, F.B., Comb, D.G., Scott, M.E., Landry, D., Vence, L.M., Perler, F.B., Benner, J., Kucera, R.B., Hirvonen, C.A., Pelletier, J.J., Paulus, H. and Xu, M.-Q. (1997). Single-column purification of free recombinant proteins using a self-cleavableaffinity tag derived from a protein splicing element. Gene. 192, 271-281.
  2. Evans, T.C., Benner, J. and Xu, M.-Q. (1998). Semisynthesis of cytotoxic proteins using a modified protein splicing element. Protein Sci.. 7, 2256-2264.
  3. Southworth, M.W., Amaya, K., Evans, J., T.C., Xu, M.-Q. and Perler, F.B. (1999). Purification of proteins fused to either the amino or carboxy terminus of the Mycobacterium xenopi gyrase A intein.. BioTechniques. 27, 110-120.
  4. Watanabe, T., Ito, Y., Yamada, T., Hashimoto, M., Sekine, S. and Tanaka, H. (1994). The role of the C-terminal domain and type III domains of chitinase A1 from Bacillus circulans WL-12 in chitin degradation. J. Bacteriol.. 176, 4465-4472.
  5. Telenti, A., Southworth, M., Alcaide, F., Daugelat, S., Jacobs, W.R. Jr. and Perler, F.B. (1997). The Mycobacterium xenopi GyrA protein splicing element: Characterization of a minimal intein. J. Bacteriol.. 179, 6378-6382.
  6. Dubendorff, J.W. and Studier, F.W. (1991). Controlling basal expression in an inducible T7 expression system by blocking the target T7 promoter with lac repressor. J. Mol. Biol.. 219, 45-59.

Protocols, Manuals & Usage

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

Application Notes

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. What vectors are included in the IMPACT kit?
  2. What is the DNA sequence of this product?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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