Expression cloning and functional characterization of human cDNA for ganglioside GM3 synthase.
AuthorsIshii A, Ohta M, Watanabe Y, Matsuda K, Ishiyama K, Sakoe K, Nakamura M, Inokuchi J, Sanai Y, Saito M
JournalJ Biol Chem
PubMed ID9822625
'Ganglioside GM3 is a major glycosphingolipid in the plasma membrane and is widely distributed in vertebrates. We describe here the isolation of a human cDNA whose protein product is responsible for the synthesis of GM3. The cloned cDNA spanned 2,359 base pairs, with an open reading frame encoding a protein ... More
Introduction of deoxyribonucleoside triphosphates into intact cells by electroporation.
Authors Sokoloski J A; Jastreboff M M; Bertino J R; Sartorelli A C; Narayanan R;
JournalAnal Biochem
PubMed ID3812972
A simple method, employing high-voltage electric discharge (electroporation), was developed to introduce phosphorylated nucleosides into the cytoplasm of viable cells. HL-60 leukemia cells permeabilized by this technique remained viable and incorporated deoxyribonucleoside triphosphates into nuclear DNA. Furthermore, DNA synthesis was depressed for at least 24 h in HL-60 cells made ... More
T cell receptor (TCR) mini-gene mRNA expression regulated by nonsense codons: a nuclear-associated translation-like mechanism
AuthorsLi S, Leonard D, Wilkinson MF
JournalJ Exp Med
PubMed ID9091590
Premature termination codons (PTCs) are known to decrease mRNA levels. Here, we report our investigation of the mechanism for this downregulation using the TCR-beta gene, which acquires PTCs as a result of programmed rearrangements that occur during normal thymic development. We found that a mini-gene version of this gene, which ... More
Expression of a human mutant monocyte chemotactic protein 3 in Pichia pastoris and characterization as an MCP-3 receptor antagonist.
AuthorsMasure S, Paemen L, Proost P, Van Damme J, Opdenakker G
JournalJ Interferon Cytokine Res
PubMed ID8590307
The cDNA encoding human monocyte chemotactic protein 3 (hMCP-3) was cloned in pHIL-S1, a vector designed for inducible secreted heterologous expression in the methylotrophic yeast Pichia pastoris. After transformation of P. pastoris by electroporation, several clones with the human MCP-3 gene integrated at the alcohol oxidase (AOX-1) locus were isolated. ... More
Short technical reports. Effects of lipopolysaccharide on transfection efficiency in eukaryotic cells.
Authors Weber M; Möller K; Welzeck M; Schorr J;
JournalBiotechniques
PubMed ID8747659
The suitability of different purification methods for preparation of plasmid DNA for transfection into eukaryotic cells was systematically investigated. The reporter plasmid, pRSVcat, was prepared using several methods, and residual impurities in the preparations were quantitated. Transfection with these preparations was performed with several cell lines (HeLa, Huh7, COS7 and ... More