Gene modification by fast-track recombineering for cellular localization and isolation of components of plant protein complexes

ABSTRACT: Genes encoding Arabidopsis subunit homologs of TFIIH kinase module were labelled with coding sequences of GFP and mCherry (green and red fluorescent proteins) using fast-track recombineering. To illustrate the applicability of recombineering for accelerating the isolation and identification of plant protein complexes, proteins associated with CDKD;2-GFP, CYCLIN-H-mCherry and DNA replication-dependent HISTONE H3.1-mCherry complexes were purified on GFP-Trap and RFP-TRAP and analysed by LC-MS/MS mass spectrometry. The results confirmed association of know TFIIH subunit homologs with CDKD;2 and CYCLIN H, and identified subunits of CAF1 (CHROMATIN ASSEMBLY FACTOR 1) and ASF1A/B histone chaperon in complex with HISTONE H3.1.

INSTRUMENT(S):

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

SUBMITTER: Sara Christina Stolze  

LAB HEAD: Hirofumi Nakagami

PROVIDER: PXD013637 | Pride | 2019-08-01

REPOSITORIES: Pride