Nawarat NantapongSutipa TanapongpipatJeffrey ColeSakol PanyimMahidol UniversityThailand National Center for Genetic Engineering and BiotechnologyUniversity of Birmingham2018-07-242018-07-242002-03-11Journal of Microbiological Methods. Vol.49, No.3 (2002), 329-334016770122-s2.0-0036008799https://repository.li.mahidol.ac.th/handle/123456789/20074An integrative plasmid containing a 1.3 kb fragment of chromosomal DNA from Enterobacter amnigenus was constructed. The Ω fragment encoding spectinomycin/streptomycin resistance was cloned into the unique BglII site of the resulting plasmid, and the interrupted fragment was transferred via plasmid pMAK705 by electroporation into E. amnigenus with a selection for spectinomycin resistance. Cointegrants were resolved to generate an E. amnigenus strain that expressed spectinomycin resistance, but grew as rapidly as the parental strain. The cloned fragment encodes a putative homologue of the proW gene of Escherichia coli that is not essential for E. amnigenus growth. The integrative plasmid is now available to introduce any heterologous DNA into the E. amnigenus chromosome, for the construction of promoter-probe vectors for the studies of gene regulation, or to construct plasmids suitable for the isolation of secretion signals. Immediate applications of this system will include the expression and secretion of crystal toxins from bacilli for the biological control of mosquito larvae infected with the bacterial host. © 2002 Elsevier Science B.V. All rights reserved.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyImmunology and MicrobiologyMedicineArticleSCOPUS10.1016/S0167-7012(01)00384-0