细胞技术

Coordinated cell type differentiation is essential for morphogenesis during Dictyostelium development. The specification of different cell types and the regulation of temporal and spatial patterns of expression of cell type-specific genes are important problems current ...

The advent of the “GFP technology” together with advances in digital imaging and microscopic techniques has revolutionized our view of the living cell. Genetically encoded fluorescent proteins have become widely used as markers in living cells. The application of these fluorescent p ...

Knockouts by homologous recombination are frequently used to investigate the function of genes in Dictyostelium and other organisms. Antisense-mediated gene silencing and RNA interference (RNAi) are convenient alternatives to reduce gene expression to different levels and ...

Dictyostelium discoideum is an exceptionally powerful system by which to study various aspects of modern cell and developmental biology. The completion of the genome sequencing project together with a high-efficiency of targeted gene disruption have enabled researchers to char ...

In the following protocols, broken cells are the starting material of all downstream purifications of functional organelles or intact subcellular membranes. The choice of the breakage method has direct and deep repercussions on the quality of subsequent steps. Breaking vegetative ...

The Dictyostelium discoideum genome has been sequenced, assembled and annotated to a high degree of reliability. The parts-list of proteins and RNA encoded by the six chromosomes can now be accessed and analyzed. One of the initial surprises was the remarkably large number of genes that are shar ...

Dictyostelium discoideum proliferates as solitary amoebae, constitutes multicellular structures called fruiting bodies, and mates to form macrocysts depending on environmental conditions. All of these processes can be easily induced in the laboratory. The amoebae are norm ...

A large number of RNA binding proteins have recently been identified that influence various human genetic disorders. However, the specific function of many of these proteins and what role they may play in a particular disease remains unclear. Identification of the substrate mRNA bound by an R ...

Efficient handling of multiple reactions is a crucial prerequisite for productive RNA differential display (DD) analysis. To identify transcriptional targets of the histone H3 Lys9-specific methyltransferase Clr4, we applied a multiformat modification of DD to compare betwe ...

Differential display technology was utilized to compare programs of gene expression in primary cultures of human skin fibroblasts from normal volunteers and patients diagnosed with melancholic depression. Polymorphic transcripts of a single gene differing by one tandem repe ...

Xbp1 is a transcriptional repressor with homology to the DNA binding domains of two cell-cycle regulatory transcription factors, Swi4 and Mbp1. Target genes bound and regulated by Xbp1, have been identified using differential display (DD). Because little was known about the regulation or ...

The glucosylation of 5-position of anthocyanin molecules is one of most important modification step, which is responsible for the stability and color hue of anthocyanin molecules. However, anthocyanin 5-O-glucosyltransferase (5-GT) is unstable and it had been difficult to purify th ...

Within the retina there is a circadian clock that controls the 24-h timing of processes such as hormone release, cell movement, and gene transcription. In an effort to better understand the molecular nature of this retinal clock, a differential display (DD) screen was performed to isolate a gene wi ...

Mammalian ovulation is a normal biological process that is initiated when a gonadotropic hormone stimulates G protein-coupled receptors in the plasma membrane of cells in ovarian follicles. This article outlines differential display (DD) protocols and associated methods that h ...

Ras signaling pathway is thought to control the expression of a subset of yet-to-be-defined genes that are crucial for cell growth and differentiation. Here we have identified by differential display a novel oncogenic Ras target gene encoding a new cytokine. Biochemical studies reveal th ...

The p53 tumor-suppressor gene is mutated in a wide range of human cancers. The ability of p53 to control passage through the cell cycle (in G1 and in G2) and to control apoptosis in response to abnormal proliferative signals and stress, including DNA damage, is considered to be important for its tumor-su ...

In order to identify molecular markers of prion disease in peripheral tissues, we used the differential display reverse-transcriptase polymerase chain reaction (DDRT-PCR) procedure to compare gene expression in spleens of infected and uninfected mice. In this study, we identified a ...

HBx and MHBst products from hepatitis B virus-DNA (HBV-DNA), which become transcriptional transactivators of cellular and viral genes, are known to play causative roles in the development of hepatocellular carcinoma (HCC). However, the biomolecular mechanism(s) for their roles in he ...

Massive amounts of DNA sequence data, generated from expressed sequence tag (EST) and genome sequencing projects, require efficient methods to link sequence databases with temporal and spatial expression profiles. To meet this need, we have developed a powerful computer program (Ge ...

Gene expression analysis by differential display (DD) is limited by the labor-intensive visual evaluation of the electrophoretic data traces. We describe a flexible method for computer-assisted ranking of expression patterns in data from DD experiments. The method is based on a pairw ...