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Publications - Cell and Metabolic Biology
Publications
Methods that enable the construction of recombinant DNA molecules are essential tools for biological research and biotechnology. Golden Gate cloning is used for assembly of multiple DNA fragments in a defined linear order in a recipient vector using a one‐pot assembly procedure. Golden Gate cloning is based on the use of a type IIS restriction enzyme for digestion of the DNA fragments and vector. Because restriction sites for the type IIS enzyme used for assembly must be present at the ends of the DNA fragments and vector but absent from all internal sequences, special care must be taken to prepare DNA fragments and the recipient vector with a structure suitable for assembly by Golden Gate cloning. In this article, protocols are presented for preparation of DNA fragments, modules, and vectors suitable for Golden Gate assembly cloning. Additional protocols are presented for assembly of defined parts in a transcription unit, as well as the stitching together of multiple transcription units into multigene constructs by the modular cloning (MoClo) pipeline.
Publications
Among the plant hormones jasmonic acid and related derivatives are known to mediate stress responses and several developmental processes. Biosynthesis, regulation, and metabolism of jasmonic acid in Arabidopsis thaliana are reviewed, including properties of mutants of jasmonate biosynthesis. The individual signalling properties of several jasmonates are described.
Publications
Transport processes between plant and fungal cells are key elements in arbuscular mycorrhiza (AM), where H+‐ATPases are considered to be involved in active uptake of nutrients from the symbiotic interface. Genes encoding H+‐ATPases were identified in the genome of Medicago truncatula and three cDNA fragments of the H+‐ATPase gene family (Mtha 1 ‐ 3) were obtained by RT‐PCR using RNA from M. truncatula mycorrhizal roots as template. While Mtha 2 and Mtha 3 appeared to be constitutively expressed in roots and unaffected by AM development, transcripts of Mtha 1 could only be detected in AM tissues and not in controls. Further analyses by RT‐PCR revealed that Mtha 1 transcripts are not detectable in shoots and phosphate availability did not affect RNA accumulation of the gene. Localization of transcripts by in situ hybridization on AM tissues showed that Mtha 1 RNA accumulates only in cells containing fungal arbuscules. This is the first report of arbuscule‐specific induced expression of a plant H+‐ATPase gene in mycorrhizal tissues.
Publications
Treatment of barley leaf segments with jasmonic acid methyl ester (JM) leads to the accumulation of a set of newly formed abundant proteins. Among them, the most abun dant protein exhibits a molecular mass of 23 kDa (JIP‐23). Here, data are presented on the occurrence and expression of the lIP‐23 genes in different cultivars of Hordeum vulgare . Southern blot analysis of 80 cultivars revealed the occurrence of 2 to 4 genes coding for JIP‐23 in all cultivars. By means of Northern blot and immunoblot analysis it is shown that some cultivars lack the ex pression of jip‐23 upon treatment of primary leaves with JM as well as upon stress performed by incubation with 1 M sorbitol solution. During germination, however, all tested cultivars ex hibited developmental expression of jip‐23 . The results are dis cussed in terms of possible functions of JIP‐23 in barley.
This page was last modified on 27 Jan 2025 .
