Abstract：In this research, genome sequencing data of Photorhabdus luminescens TT01 strains exhibited six secret genes related to the expression of the toxin complexes (Tc), including tcaZCB (plu05140515), tccAB3 (plu08050806), tccAB1 (plu41654169), tcd (plu09600971), tccC4 (plu0976) and tccC7 (plu4488), respectively. Firstly, four larger genes in TT01 chromosome named tcaZCB (6 804 bp), tccAB3 (7 419 bp), tccAB1 (10 954 bp) and tcd (51 406 bp) were rapidly fished out and constructed into vectors p15AcmtcaZCB, p15AcmtccAB3, p15Acm tccAB1, p15Acmtcd via direct cloning method. Then another two genes, tccC4 (2 891 bp) and tccC7 (2 871 bp) were gained by PCR amplification technique and formed recombinant vectors pMD18tcc4 and pMD18tcc7 by ligation with Tvector. Finally, using two different biotechnology methods, six episomal Tc genes in TT01 chromosome were successfully cloned. which would provide a positive foundation for the further study of different Tc toxins.
