Ligation-Independent Cloning for Plant Research

J.R. Wendrich, C.Y. Liao, W.A.M. van den Berg, B. De Rybel, D. Weijers

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

8 Citations (Scopus)

Abstract

Molecular cloning is a vital step in much of today’s plant biological research. Particularly, when a species is amenable to transgenic manipulation, cloning enables detailed study of gene and protein function in vivo. Therefore, accurate, consistent, and efficient cloning methods have the potential to accelerate biological research. Traditional restriction-enzyme/ligase-based strategies are often inefficient, while novel alternative methods can be less economical. We have recently optimized a method for Ligation-Independent Cloning (LIC) that is both efficient and economical. We have developed a large set of LIC-compatible plasmids for application in plant research. These include dedicated vectors for gene expression analysis, protein localization studies, and protein misexpression. We describe a detailed protocol that allows the reliable generation of plant transformation-ready constructs from PCR fragments in 2–3 days.
LanguageEnglish
Title of host publicationPlant Functional Genomics : Methods and Protocols.
EditorsJ.M. Alonso, A.N. Stepanova
Place of Publication[S.l.]
Pages421-431
DOIs
Publication statusPublished - 2015

Publication series

NameMethods in Molecular Biology
PublisherSpringer
No.1284

Fingerprint

cloning (plants)
molecular cloning
proteins
ligases
plasmids
methodology
genetically modified organisms
gene expression
enzymes
genes

Cite this

Wendrich, J. R., Liao, C. Y., van den Berg, W. A. M., De Rybel, B., & Weijers, D. (2015). Ligation-Independent Cloning for Plant Research. In J. M. Alonso, & A. N. Stepanova (Eds.), Plant Functional Genomics : Methods and Protocols. (pp. 421-431). (Methods in Molecular Biology; No. 1284). [S.l.]. https://doi.org/10.1007/978-1-4939-2444-8_21
Wendrich, J.R. ; Liao, C.Y. ; van den Berg, W.A.M. ; De Rybel, B. ; Weijers, D. / Ligation-Independent Cloning for Plant Research. Plant Functional Genomics : Methods and Protocols.. editor / J.M. Alonso ; A.N. Stepanova. [S.l.], 2015. pp. 421-431 (Methods in Molecular Biology; 1284).
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Wendrich, JR, Liao, CY, van den Berg, WAM, De Rybel, B & Weijers, D 2015, Ligation-Independent Cloning for Plant Research. in JM Alonso & AN Stepanova (eds), Plant Functional Genomics : Methods and Protocols.. Methods in Molecular Biology, no. 1284, [S.l.], pp. 421-431. https://doi.org/10.1007/978-1-4939-2444-8_21

Ligation-Independent Cloning for Plant Research. / Wendrich, J.R.; Liao, C.Y.; van den Berg, W.A.M.; De Rybel, B.; Weijers, D.

Plant Functional Genomics : Methods and Protocols.. ed. / J.M. Alonso; A.N. Stepanova. [S.l.], 2015. p. 421-431 (Methods in Molecular Biology; No. 1284).

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

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Wendrich JR, Liao CY, van den Berg WAM, De Rybel B, Weijers D. Ligation-Independent Cloning for Plant Research. In Alonso JM, Stepanova AN, editors, Plant Functional Genomics : Methods and Protocols.. [S.l.]. 2015. p. 421-431. (Methods in Molecular Biology; 1284). https://doi.org/10.1007/978-1-4939-2444-8_21