Cloning of cytolethal distending toxin A gene and construction of prokayotic expression vector
LI Lu
XU Yan
WANG Xiao-qian
YANG Mi-fang
DUAN Jun-lan
Abstract：AIM: To clone Actinobacillus actinomycetemcomitans cdtA gene and construct prokayotic expres sion vector for studying the function of CdtA protein and CDT holotoxin. METHODS: The open reading frame (ORF) of cdtA gene was cloned from the DNA of Aa. type stain ATCC 29522 by polymerase chain reaction( PCR) and cloned into pMD19 - T vector to produce the new construction pMD19 - T - cdtA. The cloned cdtA segment was cut out with two restriction enzymes and was cloned into the prokaryotic expression vector( pET15b) to produce the expression vector pET15b -cdtA. RESULTS; The prokaryotic expression vector pET15b -cdtA was successfully constructed. The cod ing region sequence of cdtA was correctly inserted into the expression vector by PCR identification and DNA sequencing analysis. Its ORF was integrated. Mutation was not detected. CONCLUSION; CdtA gene has been successfully cloned and expressed by prokaryotic expression vector(pET15b).
Keywords：cytolethal distending toxincdtA geneprokaryotic expression vector
Publication Date：2009-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 619-623 )