Dataset Information

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): Q Exactive

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

SUBMITTER: Sara Christina Stolze

LAB HEAD: Hirofumi Nakagami

PROVIDER: PXD013637 | Pride | 2019-08-01

REPOSITORIES: Pride

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			Action	DRS
	171005_CK_ZhHu_P-L4_S_01.raw	Raw
	171005_CK_ZhHu_P-L4_S_02.raw	Raw
	171005_CK_ZhHu_P-L4_S_04.raw	Raw
	171005_CK_ZhHu_P-L4_S_05.raw	Raw
	171005_CK_ZhHu_P-L4_S_06.raw	Raw

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

Hu Zhoubo Z Ghosh Ajit A Stolze Sara C SC Horváth Mihály M Bai Bing B Schaefer Sabine S Zündorf Simone S Liu Shanda S Harzen Anne A Hajheidari Mohsen M Sarnowski Tomasz J TJ Nakagami Hirofumi H Koncz Zsuzsa Z Koncz Csaba C

The Plant journal : for cell and molecular biology 20190726 2

To accelerate the isolation of plant protein complexes and study cellular localization and interaction of their components, an improved recombineering protocol is described for simple and fast site-directed modification of plant genes in bacterial artificial chromosomes (BACs). Coding sequences of fluorescent and affinity tags were inserted into genes and transferred together with flanking genomic sequences of desired size by recombination into Agrobacterium plant transformation vectors using th ...[more]

PMID: 31276249

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