Novel gateway binary vectors for rapid tripartite DNA assembly and promoter analysis with various reporters and tags in the liverwort Marchantia polymorpha

PLoS One. 2018 Oct 4;13(10):e0204964. doi: 10.1371/journal.pone.0204964. eCollection 2018.

Abstract

The liverwort Marchantia polymorpha is an emerging model species for basal lineage plant research. In this study, two Gateway cloning-compatible binary vector series, R4pMpGWB and R4L1pMpGWB, were generated to facilitate production of transgenic M. polymorpha. The R4pMpGWB series allows tripartite recombination of any promoter and any coding sequence with a specific reporter or tag. Reporters/tags for the R4pMpGWB series are GUS, ELuc(PEST), FLAG, 3×HA, 4×Myc, mRFP1, Citrine, mCitrine, ER-targeted mCitrine and nucleus-targeted mCitrine. The R4L1pMpGWB series is suitable for promoter analysis. R4L1pMpGWB vector structure is the same as that of R4pMpGWB vectors, except that the attR2 site is replaced with attL1, enabling bipartite recombination of any promoter with a reporter or tag. Reporters/tags for the R4L1pMpGWB series are GUS, G3GFP-GUS, LUC, ELuc(PEST), Citrine, mCitrine, ER-targeted mCitrine and mCitrine-NLS. Both vector series were functional in M. polymorpha cells. These vectors will facilitate the design and assembly of plasmid constructs and generation of transgenic M. polymorpha.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Artificial Gene Fusion
  • DNA, Plant / genetics*
  • Genes, Reporter / genetics*
  • Genetic Vectors / genetics*
  • Marchantia / genetics*
  • Promoter Regions, Genetic / genetics*
  • Time Factors

Substances

  • DNA, Plant

Grants and funding

This work was supported by the Ministry of Education, Culture, Sports, Science and Technology (MEXT) [KAKENHI Grant-in-Aid for Scientific Research (C) (grant Nos. 26440157 and 17K07457 to S.M), and Research Program on Hepatitis from Japan Agency for Medical Research and development, AMED, and NIBB Research Project for the Development of New Model Organisms.