[Use transposon and parDE through in vivo cloning to promote the genetic stability of plasmid pCPP430].
Plasmid pCPP430 carrying the hrp gene cluster after transformated into 308R (Pantoea agglomerans) can cause the hypersensitive response, and simultaneously induce the plant resistance to disease. It was genetically unstable in P. agglomerans. 0.8 kb parDE region of the broad-host range plasmid RK2 is responsible for plasmid partition. It can mediate plasmid maitenance of many kinds in Rhizobium meliloti, and also can promote the genetic stability of recombinant plasmid in the biocontrol bacteria P. agglomerans. In this paper, we cloned parDE into pCPP430 in vivo through transposition to promote its genetic stability. parDE was amplified by PCR, inserted into pGEM-T vector and cut out and religated to NotI-cut transposon vector pUT/mini-Tn5 Km to get a parDE containing mini-Tn5, pTnp. After conjugation between S17-1/lambda pir (pTnp) and 308R (pCPP430), parDE was cloned in vivo into plasmid pCPP430 to obtain pRTnp. It was demonstrated that the insertion of parDE in pCPP430 increased significantly the plasmid's stability in P. agglomerans.
Zhang
L
L
Institute of Biotechnology Shanxi University, Taiyuan 030006, China.
Shen
Q
Q
Zhao
L
L
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D003227:Conjugation, Genetic; D004251:DNA Transposable Elements / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D044084:Erwinia amylovora / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D042822:Genomic Instability; D020636:Pantoea / Q000235:genetics*; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C489746:ParD protein, E coli; C489748:ParD protein, Plasmid RK2; D011994:Recombinant Proteins; C075462:harpin protein, Erwinia amylovora; C078613:parD protein, Bacteria