title: Intronic T-DNA insertion in Arabidopsis NBR1 conditionally affects wild-type transcript level.
abstract: The SALK_135513 line of Arabidopsis thaliana is annotated by GenBank to have the T-DNA insertion in the fourth exon of NBR1 (At4g24690). Careful molecular analyses of the homozygous plants of SALK_135513 line indicated the place of T-DNA insertion in the fourth intron. Unexpectedly, 2 kinds of NBR1 transcripts, the wild-type and the mutated, resulting from alternative splicing events, were detected in those plants. Our findings explain the problems encountered by us with phenotypic evaluation of this line and emphasize the necessity for independent verification of the exact insertion site followed by careful expression studies when working with Arabidopsis T-DNA insertional mutants.
authors.affiliation: a Institute of Biochemistry and Biophysics ; Polish Academy of Sciences ; Warsaw , Poland.
authors.forename: Milagros Collados
authors.initials: MC
authors.lastname: Rodríguez
authors.forename: Anna
authors.initials: A
authors.lastname: Wawrzyńska
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Sirko
mesh_terms: D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics*; D002352:Carrier Proteins / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D007438:Introns / Q000235:genetics*; D016254:Mutagenesis, Insertional / Q000235:genetics*; D022821:RNA Splice Sites / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013312:Stress, Physiological
chemical_list: D029681:Arabidopsis Proteins; D002352:Carrier Proteins; D004269:DNA, Bacterial; C568852:NBR1 protein, Arabidopsis; D022821:RNA Splice Sites; D012333:RNA, Messenger; C036483:T-DNA
keywords: At4g24690; KO, knock-out; MV, methyl viologen; NBR1; NBR1, Neighbor of BRCA1; SALK_135513; T-DNA mutant; T-DNA, transferred DNA; WT – wild-type; arabidopsis; intronic insertion; qRT-PCR, quantitative RT-PCR.; sqRT-PCR, semiquantitative RT-PCR; transcription