Abstract：
:In order to develop an efficient system for deleting genomic segment in Agrobacterium tumefaciens to analyze gene functions and construct marker gene-free recombinant strains, a Cre recombinase expression plasmid was constructed by placing its encoding gene under the control of Ptet promoter and cloning into the plasmid replicable in both A. tumefaciens and E. coli. Triple recombineering was applied to efficiently construct integrative vectors which were used to introduce loxP sites and selection markers into the chromosome of A. tumefaciens. Cre recombinase could be properly induced by anhydrotetracycline in A. tumefaciens, which was revealed by the fact that kanamycin resistance gene flanked by two parallel loxP sites was excised from the genome of A. tumefaciens with virtually 100% efficiency. And what is more, an A. tumefaciens mutant carrying large-deletion (~85 kb) in genome was successfully constructed by Cre/loxP system. Here, we described the application of combination of Cre/loxP system and triple recombineering to efficiently excise genomic segment in A. tumefaciens, which also would facilitate efficient construction of multiple gene disruptions in A. tumefaciens.
journal_name
Curr Microbioljournal_title
Current microbiologyauthors
Liu Z,Xie Y,Zhang X,Hu X,Li Y,Ding X,Xia L,Hu Sdoi
10.1007/s00284-015-0977-5subject
Has Abstractpub_date
2016-04-01 00:00:00pages
465-72issue
4eissn
0343-8651issn
1432-0991pii
10.1007/s00284-015-0977-5journal_volume
72pub_type
杂志文章abstract：:In laboratory cultures of ice nucleation-active (Ice+) Erwinia herbicola isolates, it has been difficult to achieve high-level expression of ice nuclei, especially nuclei active at temperatures warmer than -5 degrees C (i.e., type 1 ice nuclei). Here we demonstrate that starvation for phosphate and exposure to low tem...
journal_title：Current microbiology
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journal_title：Current microbiology
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更新日期：2007-03-01 00:00:00
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journal_title：Current microbiology
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更新日期：2004-03-01 00:00:00
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更新日期：1999-08-01 00:00:00
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journal_title：Current microbiology
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更新日期：2001-08-01 00:00:00
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