title: [The study of optimal conditions of electroporation in Pseudomonas aeruginosa].
abstract: A P. aeruginosa strain PA68 isolated from the sputum of a patient suffering from bronchiectasis was used as the recipient strain. Optimum conditions including growth stage of the strain, electroshock voltage, concentration and preservation of competent cell were defined for the electroporation of PA68 with plasmid pSMC28. It was showed that the highest transformation efficiency was up to 1.68 x 10(3) CFU/microgram DNA under the optimum conditions in which the competent cells were collected at logarithmic growth phase (OD(540) = 0.7-0.8) and concentrated to about 10(11) cells/ml, the mixture of the competent cells and plasmid pSMC28 was eletroporated at 2.6 kV. With this optimal condition, Mu transponson complexes have been successfully transformed into P. aeruginosa strain PA68 and the obtained efficiency was 2.47 x 10(4) CFU/microgram DNA. This is the first time to electroporate Mu transposon complexes into Pseudomonas spp. The artificial Mu transposons could integrate into bacterial genomes at a single site randomly. Then the phenotype change was the result of the gene inactivation caused by Mu transposon insertion. That will be very helpful for the study of genomic function of Pseudomonas spp.
authors.affiliation: College of Life Sciences, Nankai University, Tianjin 300071, China. zhiyingshan@163.com
authors.forename: Zhi-Ying
authors.initials: ZY
authors.lastname: Shan
authors.forename: Hai-Jin
authors.initials: HJ
authors.lastname: Xu
authors.forename: Xing-Qi
authors.initials: XQ
authors.lastname: Shi
authors.forename: Zhou
authors.initials: Z
authors.lastname: Nie
authors.forename: Yan
authors.initials: Y
authors.lastname: Yu
authors.forename: Xiu-Ming
authors.initials: XM
authors.lastname: Zhang
authors.forename: Yan-Ling
authors.initials: YL
authors.lastname: Bai
authors.forename: Ming-Qiang
authors.initials: MQ
authors.lastname: Qiao
authors.forename: Cai-Chang
authors.initials: CC
authors.lastname: Gao
mesh_terms: D004251:DNA Transposable Elements; D018274:Electroporation; D006801:Humans; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000254:growth & development; D014169:Transformation, Bacterial; D019895:Transposases / Q000502:physiology
chemical_list: D004251:DNA Transposable Elements; C114697:mu transposase; D019895:Transposases