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TargeTron vector pJIR750ai is a circularized vector that 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 to 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, TA0100 User Guide (1.1 MB PDF).
TargeTron is a registered trademark of InGex, LLC.
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TargeTron® Gene Knockout System
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