Retrovirus promoter-trap vector to induce lacZ gene fusions in mammalian cells

LacZ-promoter fusions: the effect of growth.

A lacZ-hygromycin fusion gene and its use in a gene trap vector for marking embryonic stem cells.

The use of gene trap vectors in embryonic stem (ES) cells and transgenic mice has proved to be a powerful method for identifying new genes involved in mammalian development (1—4). Generally, they have a lacZ marker gene downstream of a splice acceptor site and rely on integration into introns for their expression. Not only can insertion of these vectors cause disruption of the gene into which t...

Alpha complementation of LacZ in mammalian cells.

The bacterial lactose converting enzyme beta-galactosidase (β-gal) and its gene (LacZ) have been studied for many years (1 and references therein) and are among the most utilized tools in molecular biology. The LacZ product, a polypeptide of 1029 amino acids, gives rise to the functional enzyme after tetramerization (2 and references therein) and is easily detected by chromogenic substrates eit...

Vector Integration Sites Identification for Gene-Trap Screening in Mammalian Haploid Cells

Forward genetic screens using retroviral (or transposon) gene-trap vectors in a haploid genome revolutionized the investigation of molecular networks in mammals. However, the sequencing data generated by Phenotypic interrogation followed by Tag sequencing (PhiT-seq) were not well characterized. The analysis of human and mouse haploid screens allowed us to describe PhiT-seq data and to define qu...

An avian sarcoma/leukosis virus-based gene trap vector for mammalian cells.

RCASBP-M2C is a retroviral vector derived from an avian sarcoma/leukosis virus which has been modified so that it uses the envelope gene from an amphotropic murine leukemia virus (E. V. Barsov and S. H. Hughes, J. Virol. 70:3922-3929, 1996). The vector replicates efficiently in avian cells and infects, but does not replicate in, mammalian cells. This makes the vector useful for gene delivery, m...