Conversion of BAC Clones into Binary BAC (BIBAC) Vectors and Their Delivery into Basidiomycete Fungal Cells Using Agrobacterium tumefaciens

Handle URI:
http://hdl.handle.net/10754/622139
Title:
Conversion of BAC Clones into Binary BAC (BIBAC) Vectors and Their Delivery into Basidiomycete Fungal Cells Using Agrobacterium tumefaciens
Authors:
Ali, Shawkat ( 0000-0003-3282-3186 ) ; Bakkeren, Guus
Abstract:
The genetic transformation of certain organisms, required for gene function analysis or complementation, is often not very efficient, especially when dealing with large gene constructs or genomic fragments. We have adapted the natural DNA transfer mechanism from the soil pathogenic bacterium Agrobacterium tumefaciens, to deliver intact large DNA constructs to basidiomycete fungi of the genus Ustilago where they stably integrated into their genome. To this end, Bacterial Artificial Chromosome (BAC) clones containing large fungal genomic DNA fragments were converted via a Lambda phage-based recombineering step to Agrobacterium transfer-competent binary vectors (BIBACs) with a Ustilago-specific selection marker. The fungal genomic DNA fragment was subsequently successfully delivered as T-DNA through Agrobacterium-mediated transformation into Ustilago species where an intact copy stably integrated into the genome. By modifying the recombineering vector, this method can theoretically be adapted for many different fungi.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division; Center for Desert Agriculture
Citation:
Ali S, Bakkeren G (2014) Conversion of BAC Clones into Binary BAC (BIBAC) Vectors and Their Delivery into Basidiomycete Fungal Cells Using Agrobacterium tumefaciens. Bacterial Artificial Chromosomes: 199–215. Available: http://dx.doi.org/10.1007/978-1-4939-1652-8_9.
Publisher:
Springer Science + Business Media
Journal:
Bacterial Artificial Chromosomes
Issue Date:
19-Sep-2014
DOI:
10.1007/978-1-4939-1652-8_9
Type:
Book Chapter
ISSN:
1064-3745; 1940-6029
Appears in Collections:
Center for Desert Agriculture; Book Chapters; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorAli, Shawkaten
dc.contributor.authorBakkeren, Guusen
dc.date.accessioned2017-01-02T08:10:21Z-
dc.date.available2017-01-02T08:10:21Z-
dc.date.issued2014-09-19en
dc.identifier.citationAli S, Bakkeren G (2014) Conversion of BAC Clones into Binary BAC (BIBAC) Vectors and Their Delivery into Basidiomycete Fungal Cells Using Agrobacterium tumefaciens. Bacterial Artificial Chromosomes: 199–215. Available: http://dx.doi.org/10.1007/978-1-4939-1652-8_9.en
dc.identifier.issn1064-3745en
dc.identifier.issn1940-6029en
dc.identifier.doi10.1007/978-1-4939-1652-8_9en
dc.identifier.urihttp://hdl.handle.net/10754/622139-
dc.description.abstractThe genetic transformation of certain organisms, required for gene function analysis or complementation, is often not very efficient, especially when dealing with large gene constructs or genomic fragments. We have adapted the natural DNA transfer mechanism from the soil pathogenic bacterium Agrobacterium tumefaciens, to deliver intact large DNA constructs to basidiomycete fungi of the genus Ustilago where they stably integrated into their genome. To this end, Bacterial Artificial Chromosome (BAC) clones containing large fungal genomic DNA fragments were converted via a Lambda phage-based recombineering step to Agrobacterium transfer-competent binary vectors (BIBACs) with a Ustilago-specific selection marker. The fungal genomic DNA fragment was subsequently successfully delivered as T-DNA through Agrobacterium-mediated transformation into Ustilago species where an intact copy stably integrated into the genome. By modifying the recombineering vector, this method can theoretically be adapted for many different fungi.en
dc.publisherSpringer Science + Business Mediaen
dc.subjectAgrobacterium-mediated transformationen
dc.subjectBacterial artificial chromosomeen
dc.subjectBasidiomyceteen
dc.subjectBinary vectoren
dc.subjectRecombineeringen
dc.subjectUstilagoen
dc.titleConversion of BAC Clones into Binary BAC (BIBAC) Vectors and Their Delivery into Basidiomycete Fungal Cells Using Agrobacterium tumefaciensen
dc.typeBook Chapteren
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.contributor.departmentCenter for Desert Agricultureen
dc.identifier.journalBacterial Artificial Chromosomesen
dc.contributor.institutionAgriculture and Agri-Food Canada, Pacific Agri-Food Research Centre, Summerland, BC, Canadaen
dc.contributor.institutionDepartment of Botany, University of British Columbia, Vancouver, BC, Canadaen
kaust.authorAli, Shawkaten
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