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T7701

Sigma-Aldrich

TargeTron Vector pJIR750ai

Expression Vector for Bacterial Gene Knockout

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storage temp.

−20°C

General description

TargeTron vector pJIR750ai is a 10,262 bp expression vector intended for use with the TargeTron Gene Knockout System, Catalog Number TA0100. This circularized vector can be used for targeted gene knockouts in gram-positive bacteria such as Clostridium perfringens type A.1 Expression of the group II intron RNA is under the control of the β-2 toxin gene promoter, cpb2.1 As supplied, the pJIR750ai vector is re-targeted to the α-toxin (plc) gene. α-Toxin, a phospholipase C, is an extracellular toxin produced by all C. perfringens isolates.1 For validation in a specific C. perfringens strain, this vector can be used directly to knockout plc without any further modifications using the enclosed protocol. In order to re-target this vector to knockout other genes, the plc specific IBS-EBS fragment (350 bp) between the Hind III and BsrG I sites can be cut out and replaced with another gene specific fragment. For re-targeting protocols, refer to the TargeTron Gene Knockout System User Guide, Catalog Number TA0100, at sigma-aldrich.com.

Legal Information

TargeTron is a trademark of InGex, LLC

Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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Courtney L Frazier et al.
Applied and environmental microbiology, 69(2), 1121-1128 (2003-02-07)
Despite their commercial importance, there are relatively few facile methods for genomic manipulation of the lactic acid bacteria. Here, the lactococcal group II intron, Ll.ltrB, was targeted to insert efficiently into genes encoding malate decarboxylase (mleS) and tetracycline resistance (tetM)
Jun Yao et al.
RNA (New York, N.Y.), 12(7), 1271-1281 (2006-06-03)
We show that a targetron based on the Lactococcus lactis Ll.LtrB group II intron can be used for efficient chromosomal gene disruption in the human pathogen Staphylococcus aureus. Targetrons expressed from derivatives of vector pCN37, which uses a cadmium-inducible promoter
Yue Chen et al.
Applied and environmental microbiology, 71(11), 7542-7547 (2005-11-05)
In developing Clostridium perfringens as a safe vaccine vector, the alpha toxin gene (plc) in the bacterial chromosome must be permanently inactivated. Disrupting genes in C. perfringens by traditional mutagenesis methods is very difficult. Therefore, we developed a new strategy
Jones, J.P., et al.
Molecular Cancer Therapeutics, 11, 687-694 (2005)
Jin Zhong et al.
Nucleic acids research, 31(6), 1656-1664 (2003-03-11)
Mobile group II introns have been used to develop a novel class of gene targeting vectors, targetrons, which employ base pairing for DNA target recognition and can thus be programmed to insert into any desired target DNA. Here, we have

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