Molecular cloning and analysis of four potato tuber mRNAs

Stiekema, Willem J.; Heidekamp, F.; Dirkse, W. G.; Beckum, J van; Haan, Peter; Bosch, C T; Louwerse, Jeanine

doi:10.1007/bf00027383

Cited by 208 publications

(181 citation statements)

References 51 publications

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“…In addition to DNA sequence information, the presence of suitable Asel and Taql sites in the coding region of the gene was a prerequisite. The patatin B2 gene (STPATB2, EMBL Data Library accession number X13178; Stiekema et al, 1988) was found to fulfil these criteria. The TDF was predicted to yield a fragment of 570 bp when selective base exten- sions AA (Asel-end) and AC (Taql-end) were used on the 3' ends of the primers.…”

Section: Analysis Of Gene Expression Using Cdna-aflp and Northern Anamentioning

confidence: 99%

Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development

Bachem¹,

Hoeven²,

et al. 1996

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SummaryUsing a highly synchronous in vitro tuberization system, in combination with an amplified restriction fragment polymorphism (AFLpt)-derived technique for RNA fingerprinting (cDNA-AFLPt), transcriptional changes at and around the time point of potato tuberization have been analyzed. The targeted expression analysis of a specific transcript coding for the major potato storage protein, patatin and a second transcript, coding for ADP-glucose pyrophosphorylase, a key gene in the starch biosynthetic pathway is described. This paper confirms that kinetics of expression revealed by cDNA-AFLP analysis are comparable to those found in Northern analysis. Furthermore, this paper reports the isolation and analysis of two tuberspecific transcript-derived fragments (TDFs) coding for the lipoxygenase enzyme, which are differentially induced around the time point of tuber formation. Analysis of the two Iox TDFs demonstrates that it is possible to dissect the expression modalities of individual transcripts, not independently detectable by Northern analysis. Finally, it is shown that using cDNA-AFLP, rapid and simple verification of band identity may be achieved. The results indicate that cDNA-AFLP is a broadly applicable technology for identifying developmentally regulated genes.

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Section: Analysis Of Gene Expression Using Cdna-aflp and Northern Anamentioning

confidence: 99%

Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development

Bachem¹,

Hoeven²,

et al. 1996

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“…Plant defensins have initially been identified in seeds , and recently been reported in other plant organs and tissues, e.g., leaves, flower, fruit, pods and tubers (Stiekema et al 1988;Chiang and Hadwiger 1991;Meyer et al 1996;Urdangarin et al 2000). They play a significant role in host defense.…”

Section: Introductionmentioning

confidence: 99%

Antifungal activity of a recombinant defensin CADEF1 produced by Escherichia coli

2009

World J Microbiol Biotechnol

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The defensin CADEF1 has been known to be uptranscripted in the pepper Capsicum annuum L., upon bacterial attack, abiotic elicitors and environmental stresses. However, the native form of CADEF1 has not been purified from the pepper, and its activity against fungi has been uncertain. In this study, the full-length cDNA of CADEF1 was obtained by fragment piecing-together method. Thereafter, its mature peptide coding sequence was inserted into bacterial expression vector pET28a(?), and the recombinant vector was transformed into Escherichia coli BL21 (DE3). Based on SDS-PAGE and Western blotting analysis, the recombinant CADEF1 was produced as a 5.6 kDa protein in the form of inclusion body in E. coli BL21 (DE3) and the expression level accounted for 15% of the bacterial total protein. Further processing through protein refolding and purification, converted the inclusion body type of CADEF1 into active form with the yield about 13 lg/ml of culture. The refolded CADEF1 significantly showed the activity against growth of the fungal pathogen Verticillium dahliae on average by 67.9% in vitro assay. These results verified the novel activity of CADEF1, which might benefit in prevention of the fungus-related disease in agricultural production.

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“…cDNA probes were prepared from total RNA isolated from vegetative mycelium and knobs, grown under identical conditions to those used in the construction of the cDNA libraries. Total RNA was extracted according to Stiekema et al (1988) and purified further on an RNeasy plant mini kit spin column (Qiagen). To obtain the required amounts of RNA from limited amounts of starting material, two rounds of antisense RNA (aRNA) amplification were performed using the MessageAmp kit (Ambion).…”

Section: Methodsmentioning

confidence: 99%

Comparison of gene expression in trap cells and vegetative hyphae of the nematophagous fungus Monacrosporium haptotylum

et al. 2005

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Nematode-trapping fungi enter the parasitic stage by developing specific morphological structures called traps. The global patterns of gene expression in traps and mycelium of the fungus Monacrosporium haptotylum were compared. The trap of this fungus is a unicellular spherical structure called the knob, which develops on the apex of a hyphal branch. RNA was isolated from knobs and mycelium and hybridized to a cDNA array containing probes of 2822 EST clones of M. haptotylum. Despite the fact that the knobs and mycelium were grown in the same medium, there were substantial differences in the patterns of genes expressed in the two cell types. In total, 23?3 % (657 of 2822) of the putative genes were differentially expressed in knobs versus mycelium. Several of these genes displayed sequence similarities to genes known to be involved in regulating morphogenesis and cell polarity in fungi. Among them were several putative homologues for small GTPases, such as rho1, rac1 and ras1, and a rho GDP dissociation inhibitor (rdi1). Several homologues to genes involved in stress response, protein synthesis and protein degradation, transcription, and carbon metabolism were also differentially expressed. In the last category, a glycogen phosphorylase (gph1) gene homologue, one of the most upregulated genes in the knobs as compared to mycelium, was characterized. A number of the genes that were differentially expressed in trap cells are also known to be regulated during the development of infection structures in plant-pathogenic fungi. Among them, a gas1 (mas3) gene homologue (designated gks1), which is specifically expressed in appressoria of the rice blast fungus, was characterized.

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Molecular cloning and analysis of four potato tuber mRNAs

Cited by 208 publications

References 51 publications

Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development

Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development

Antifungal activity of a recombinant defensin CADEF1 produced by Escherichia coli

Comparison of gene expression in trap cells and vegetative hyphae of the nematophagous fungus Monacrosporium haptotylum

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