细胞技术

The gene trap approach, represented in Fig. 1, is based on the use of mouse embryonic stem (ES) cells and a class of vectors containing a splice-acceptor site upstream of the reporter and resistance genes, β-galactosidase (β-gal, lacZ) and neomycin resistance (neo), respectively. Integration of ...

Immunologic reagents such as antibodies can be valuable for identifying and isolating cells with particular characteristics. While antibodies derived from immunized animals are the most common immunologic reagents used for this purpose, “antibodies” selected by phage displ ...

Phage-mediated gene transfer offers an alternative method of introducing genes into specific cell types, including cell lines (1–4) and primary cell cultures (see Fig. 1). Recent studies demonstrate that filamentous bacteriophages can be engineered to transfer genes to mammalian c ...

Knowledge of the molecular and cellular events characterizing osteoblast development is growing as new markers, including important classes of regulatory molecules such as transcription factors (e.g., Cbfa-1 ), are elucidated. Nevertheless, a paucity of definitive and specific ...

During embryonic development, primitive ectoderm forms three primary germ layers, the mesoderm, the ectoderm, and the endoderm. These germ layers interact forming all the tissues and organs of the developing embryo. The influences controlling the transition of ectoderm to visceral a ...

Cellular gene expression changes during ontogenetic development of cell physiological activation-inhibtion and differentiation. Classical molecular assays like Southern, Northern, or Western blotting display only a few genes at once. The analysis of complex alterations ...

Expression studies performed on a genome scale have become very popular and provide an important link between the sequence and the function of a specific gene, thereby constituting the first step towards the elucidation of the function of specific genes. By correlating the modulation of gene ...

The control of gene function by the modulation of mRNA translation, stability, or protein activity has important implications for the treatment of disease, as well as in research programs designed to study the biological role of proteins. For instance, preventing protein function in spec ...

A first step in defining the function of a novel gene is to determine its interactions with other gene products in an appropriate context; that is, because proteins make specific interactions with other proteins as part of functional assemblies, an appropriate way to examine the function of the p ...

The two-hybrid system (THS) (1) is a molecular genetic screen that detects protein-protein interactions. The protein specified by the yeast GAL4 gene activates the transcription of genes involved in galactose metabolism. It has two functional domains, a DNA binding domain, Gal4BD, and a tr ...

Dozens of genomes will be partly or completely sequenced over the next few years. In the post genomic area, we are now facing the challenge of functionally characterizing thousands of genes generated by the genome projects. Selective enrichment of clones encoding a desired gene product and ra ...

The molecular mechanisms underlying self-renewal of pluripotent embryonic stem (ES) cells is still poorly understood. Deciphering these mechanisms is of prime importance for at least two reasons: (1) ES cells derive from, and are closely related to, the pluripotent stem cells of the blast ...

The early phase of murine embryonic stem (ES) cell differentiation is accompanied with the expression of proteins that are key players in this phenomenon. The function of some these proteins can be analyzed by using the classical “knock-out gene technology”, that consists of the inactivati ...

Little is known about the molecular mechanisms controlling the earliest cellular differentiation events of the mammalian embryo. Pluripotent embryonic stem (ES) cell lines, derived from cells of the inner cell mass of the blastocyst, are an important experimental system that can be used ...

Gap junctions are specialized cell-cell junctions that directly link the cytoplasm of neighboring cells. They mediate the direct transfer of low molecular weight (

Mammalian development occurs in the hypoxic environment of the uterus (1,2). Initially, the limited oxygen available can adequately diffuse to all the cells of the growing conceptus. However, with continued growth, diffusion becomes less efficient and results in a “physiologic hypox ...

With the attractive potential therapeutic uses of both endothelial and hematopoietic cells in medicine, much attention has been focused upon the differentiation of embryonic stem (ES) cells into mature cells of these lineages. In this chapter, we present a culture system that has success ...

Numerous causative genes involved in human cancers, such as leukemias and lymphomas, have been identified (1). Considerable evidence has accumulated showing that disruptions of these genes can affect hematopoietic cell differentiation and growth. However, it remains largely un ...

There are two key methods which yield information about the cellular function of a protein. One can introduce into cells or whole organisms a form of the protein that is constitutively active, and which therefore functions independent of the normal regulatory mechanisms. This technique has ...

Members of the Ras superfamily of small GTPases (p21) are involved in the regulation of a large variety of key cellular processes, including cell differentiation and proliferation, membrane trafficking, and nuclear import and export. Based on sequence homology, this superfamily can be d ...