Gene inactivation in Arabidopsis thaliana is not accompanied by an accumulation of repeat-induced point mutations.
Chromosomal integration of multicopy transgene inserts in higher plants is often followed by loss of expression. We have analysed whether this inactivation can trigger repeat-induced point mutations (RIP) as has been observed in Neurospora crassa. We have previously characterized transgenic lines of Arabidopsis thaliana containing the hygromycin phosphotransferase (hpt) gene either as a unique sequence in plants expressing the gene, or as multimeric, closely linked repeats in clones that were resistant to hygromycin directly after transformation but exhibited gene inactivation in the subsequent generation. At the sequence level, we have determined the mutation frequencies in the promoter and coding regions of active and inactive copies of transgene inserts after passage through three sexual generations. No RIP-like mutations were found in inactivated genes. Comparison of our data with those from Neurospora suggest that sequence divergence within plant repetitive DNA is either much slower than in Neurospora or is generated by a different mechanism.
Mittelsten Scheid
O
O
Friedrich Miescher Institute, Basel, Switzerland.
Afsar
K
K
Paszkowski
J
J
D017360:Arabidopsis / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D017343:Genes, Plant / Q000235:genetics*; D008040:Genetic Linkage; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D017354:Point Mutation; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase