Method for Biolistic Site-Specific Integration in Plants Catalyzed by Bxb1 Integrase

Ruyu Li; Zhiguo Han; Lili Hou; Gurminder Kaur; Qian Yin; David W Ow

doi:10.1007/978-1-4939-4931-1_2

Method for Biolistic Site-Specific Integration in Plants Catalyzed by Bxb1 Integrase

Methods Mol Biol. 2016:1469:15-30. doi: 10.1007/978-1-4939-4931-1_2.

Authors

Ruyu Li¹, Zhiguo Han¹, Lili Hou^{1

2}, Gurminder Kaur^{1

2}, Qian Yin^{1

2}, David W Ow³

Affiliations

¹ Plant Gene Engineering Center, South China Botanical Garden, Chinese Academy of Sciences, 723 Xingke Road, Guangzhou, 510650, China.
² University of Chinese Academy of Sciences, Beijing, China.
³ Plant Gene Engineering Center, South China Botanical Garden, Chinese Academy of Sciences, 723 Xingke Road, Guangzhou, 510650, China. dow@scbg.ac.cn.

PMID: 27557683
DOI: 10.1007/978-1-4939-4931-1_2

Abstract

Crop improvement is a never ending process. With a transgenesis approach, it is not inconceivable to envision a continuous addition of new transgenes to existing cultivars. Previously, we described a recombinase-directed gene stacking method in tobacco (Hou et al., Mol Plant 7:1756-1765, 2014). Being able to stack DNA to a previous location ensures that the number of genetic loci does not increase with each new round of transgene addition. Whereas the previous demonstration was conducted through polyethylene glycol to mediate uptake of DNA into tobacco protoplasts, we now describe protocols for using biolistic transformation to stack DNA in tobacco and rice.

Keywords: Gene stacking; Gene targeting; Recombinase; Site-specific recombination; Transformation; Transgene expression.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biolistics / methods*
Integrases / genetics
Nicotiana / genetics*
Oryza / genetics*
Plants, Genetically Modified / genetics*
Polymerase Chain Reaction
Recombination, Genetic
Transgenes*

Substances

Integrases