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T A Richmond

Publications and source records attributed to T A Richmond.

6 recordsLinked to original sources
PubMed2006

Ultra-high resolution array painting facilitates breakpoint sequencing.

OBJECTIVE: To describe a considerably advanced method of array painting, which allows the rapid, ultra-high resolution mapping of translocation breakpoints such that rearrangement junction fragments can be amplified directly and sequenced. METHOD: Ultra-high resolution array painting involves the hybridisation of probes generated by the amplification of small numbers of flow-sorted derivative chromosomes to oligonucleotide arrays designed to tile breakpoint regions at extremely high resolution. RESULTS AND DISCUSSION: How ultra-high resolution array painting of four balanced translocation cases rapidly and efficiently maps breakpoints to a point where junction fragments can be amplified easily and sequenced is demonstrated. With this new development, breakpoints can be mapped using just two array experiments: the first using whole-genome array painting to tiling resolution large insert clone arrays, the second using ultra-high-resolution oligonucleotide arrays targeted to the breakpoint regions. In this way, breakpoints can be mapped and then sequenced in a few weeks.

Adult↗
PubMed2001

Integrative approaches to determining Csl function.

While there is an ever-increasing amount of information regarding cellulose synthase catalytic subunits (CesA) and their role in the formation of the cell wall, the remainder of the enzymes that synthesize structural cell wall polysaccharides are unknown. The completion of the Arabidopsis genome and the wealth of the sequence information from other plant genome projects provide a rich resource for determining the identity of these enzymes. Arabidopsis contains six families of genes related to cellulose synthase, the cellulose synthase-like (Csl) genes. Our laboratory is taking a multidisciplinary approach to determine the function of the Csl genes, incorporating genomic, genetic and biochemical data. Information from expressed sequence tag (EST) projects has revealed the presence of Csl genes in all plant species with a significant number of ESTs. Certain Csl families appear to be missing from some species. For example, no examples of CslG ESTs have been found in rice or maize. Microarray data and reporter constructs are being used to determine the expression pattern of the CesA and Csl genes in Arabidopsis. Mutations and insertion events have been identified in a majority of the genes in the Arabidopsis CesA superfamily and are being characterized by phenotypic and biochemical analysis. While we cannot yet link the function of any of the Csl genes to their respective products, the expression and localization of these genes is consistent with the expected expression pattern of polysaccharide synthases that contribute to the primary cell wall.

Arabidopsis↗
PubMed2000

Engineered metal binding sites on green fluorescence protein.

The ability to assay a variety of metals by noninvasive methods has applications in both biomedical and environmental research. Green fluorescent protein (GFP) is a protein isolated from coelenterates that exhibits spontaneous fluorescence. GFP does not require any exogenous cofactors for fluorescence, and can be easily appended to other proteins at the DNA level, producing a fluorescence-labeled target protein in vivo. Metals in close proximity to chromophores are known to quench fluorescence in a distance-dependent fashion. Potential metal binding sites on the surface of GFP have been identified and mutant proteins have been designed, created, and characterized. These metal-binding mutants of GFP exhibit fluorescence quenching at lower transition metal ion concentrations than those of the wild-type protein. These GFP mutants represent a new class of protein-based metal sensors.

Binding Sites↗
PubMed1999

A defect in beta-oxidation causes abnormal inflorescence development in Arabidopsis.

The abnormal inflorescence meristem1 (aim1) mutation affects inflorescence and floral development in Arabidopsis. After the transition to reproductive growth, the aim1 inflorescence meristem becomes disorganized, producing abnormal floral meristems and resulting in plants with severely reduced fertility. The derived amino acid sequence of AIM1 shows extensive similarity to the cucumber multifunctional protein involved in beta-oxidation of fatty acids, which possesses l-3-hydroxyacyl-CoA hydrolyase, l-3-hydroxyacyl-dehydrogenase, d-3-hydroxyacyl-CoA epimerase, and Delta(3), Delta(2)-enoyl-CoA isomerase activities. A defect in beta-oxidation has been confirmed by demonstrating the resistance of the aim1 mutant to 2,4-diphenoxybutyric acid, which is converted to the herbicide 2,4-D by the beta-oxidation pathway. In addition, the loss of AIM1 alters the fatty acid composition of the mature adult plant.

Amino Acid Sequence↗
PubMed1994

The fate of inflorescence meristems is controlled by developing fruits in Arabidopsis.

The relationship between fruit development and the proliferative capacities of inflorescence meristems has been examined in Arabidopsis thaliana. In the wild-type Landsberg erecta (Ler) line, flower production ceases coordinately on all inflorescence branches by a process we have designated global proliferative arrest (GPA). Morphological studies indicate that GPA involves a cessation of proliferative activity at the meristems, but a retention of the structural characteristics of the proliferating meristems. GPA does not occur in the male-sterile (ms1-1) line, nor in wild-type Ler when fruits are surgically removed. In these cases, inflorescence meristems continue to proliferate, ultimately terminating by a different process, designated terminal differentiation, in which disruptions in patterning at the apex are followed by the loss of the inflorescence meristem. We present an argument that GPA is mediated by a specific communication system between inflorescence meristems and developing fruits. Analysis of reduced-fertility mutants provided evidence that GPA is dependent on seed development specifically. Mutations conferring hormone deficiency or insensitivity did not disrupt the correlative interactions leading to GPA.

Arabidopsis↗