Dissociation (Ds) constructs, mapped Ds launch pads and a transiently-expressed transposase system suitable for localized insertional mutagenesis in rice

Narayana M Upadhyaya; Qian-Hao Zhu; KEFAN Peng; Song J Kim; Elizabeth S Dennis; Xue Zhou; Andrew Eamens; Mohammad S Hoque; Kerrie Ramm; Ramannee Shivakkumar; Kathryn F Smith; Shu-Ting Pan; SUZHI LI

doi:10.1007/s00122-006-0235-0

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Dissociation (Ds) constructs, mapped Ds launch pads and a transiently-expressed transposase system suitable for localized insertional mutagenesis in rice

Journal article

Peer reviewed

Dissociation (Ds) constructs, mapped Ds launch pads and a transiently-expressed transposase system suitable for localized insertional mutagenesis in rice

Narayana M Upadhyaya, Qian-Hao Zhu, KEFAN Peng, Song J Kim, Elizabeth S Dennis, Xue Zhou, Andrew Eamens, Mohammad S Hoque, Kerrie Ramm, Ramannee Shivakkumar, …

Theoretical and Applied Genetics, Vol.112(7), pp.1326-1341

2006

DOI: https://doi.org/10.1007/s00122-006-0235-0

PMID: 16505997

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Abstract

Insertion Line

Callus Line

Vector Backbone

Gene Trap

Double Transformants

We have developed a transiently-expressed transposase (TET)-mediated Dissociation (Ds) inser-tional mutagenesis system for generating stable insertion lines in rice which will allow localized mutagenesis of a chromosomal region. In this system, a Ds containing T-DNA construct was used to produce Ds launch pad lines. Callus tissues, from single-copy Ds/T-DNA lines, were then transiently infected with Agrobacterium harbouring an immobile Ac (iAc) construct, also containing a green fluorescent protein gene (sgfpS65T) as the visual marker. We have regenerated stable Ds insertion lines at a frequency of 9–13% using selection for Ds excision and GFP counter selection against iAc and nearly half of them were unique insertion lines. Double transformants (iAc/Ds) were also obtained and their progeny yielded $10% stable insertion lines following excision and visual marker screening with 50% redundancy. In general, more than 50% of the Ds reinsertions were within 1 cM of the launch pad.

Details

Title: Dissociation (Ds) constructs, mapped Ds launch pads and a transiently-expressed transposase system suitable for localized insertional mutagenesis in rice
Authors: Narayana M Upadhyaya (Author) - CSIRO Plant Industry
Qian-Hao Zhu (Author) - CSIRO Plant Industry
KEFAN Peng (Author) - CSIRO Plant Industry
Song J Kim (Author) - CSIRO Plant Industry
Elizabeth S Dennis (Author) - CSIRO Plant Industry
Xue Zhou (Author) - CSIRO Plant Industry
Andrew Eamens (Author) - CSIRO Plant Industry
Mohammad S Hoque (Author) - CSIRO Plant Industry
Kerrie Ramm (Author) - CSIRO Plant Industry
Ramannee Shivakkumar (Author) - CSIRO Plant Industry
Kathryn F Smith (Author) - CSIRO Plant Industry
Shu-Ting Pan (Author) - CSIRO Plant Industry
SUZHI LI (Author) - CSIRO Plant Industry
Publication details: Theoretical and Applied Genetics, Vol.112(7), pp.1326-1341
Publisher: Springer
DOI: 10.1007/s00122-006-0235-0
ISSN: 1432-2242
PMID: 16505997
Organisation Unit: School of Health and Behavioural Sciences - Legacy; University of the Sunshine Coast, Queensland; School of Health - Biomedicine
Language: English
Record Identifier: 99656588902621
Output Type: Journal article

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Web Of Science research areas: Agronomy; Genetics & Heredity; Horticulture; Plant Sciences

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