Hygromycin B phosphotransferase as a selectable marker for DNA transfer experiments with higher eucaryotic cells.

Blöchlinger, Karen; Diggelmann, Heidi

doi:10.1128/mcb.4.12.2929

Cited by 254 publications

(96 citation statements)

References 12 publications

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“…A bovine papilloma virus-based expression vector containing human insulin receptor cDNA (27,28) was transfected by electroporation into clone HR15. Selection of stably transfected cells was achieved by cotransfection of a hygromycin B phosphotransferase expression plasmid (29) followed by culture in the presence of hygromycin B (0.3 mg/ml) (Sigma) for 10 days.…”

Section: Methodsmentioning

confidence: 99%

Targeted inactivation of the insulin receptor gene in mouse 3T3-L1 fibroblasts via homologous recombination.

Accili

Taylor

1991

Proc. Natl. Acad. Sci. U.S.A.

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To study the role of the insulin receptor in determining adipocyte differentiation of the mouse cell line 3T3-L1, we have introduced a mutation that inactivates the insulin receptor gene by homologous recombination. In two independent clones, inactivation of one allele of the insulin receptor gene was associated with a 50-70% reduction in the number, of insulin receptors. In addition, both clones were markedly impaired in their ability to differentiate into adipocytes. The defect in adipocyte-specific differentiation was corrected by expression of transfected human insulin receptor cDNA. These data suggest that the insulin receptor may play an important role in promoting differentiation of 3T3-L1 cells into adipocytes in vitro.

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Section: Methodsmentioning

confidence: 99%

Targeted inactivation of the insulin receptor gene in mouse 3T3-L1 fibroblasts via homologous recombination.

Accili

Taylor

1991

Proc. Natl. Acad. Sci. U.S.A.

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“…For stable transfection, HeLa rtTA HR5 (BD Biosciences Clontech, France) cells were co-transfected with 6 µg tTA/CPEB1-∆5-long plasmid and 0.6 µg of hygromycin-resistant pY3 plasmid DNA (Blochlinger and Diggelmann, 1984) per 100 mm diameter dish using lipofectamine 2000 (Invitrogen, France). The selection of stable clones was achieved as described previously (Audibert et al, 2002).…”

Section: Cell Culturementioning

confidence: 99%

The translational regulator CPEB1 provides a link between dcp1 bodies and stress granules

Wilczynska

Aigueperse

Kress

et al. 2005

Journal of Cell Science

273

291

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The cytoplasmic polyadenylation element-binding protein (CPEB) has been characterized in Xenopus laevis as a translational regulator. During the early development, it behaves first as an inhibitor and later as an activator of translation. In mammals, its closest homologue is CPEB1 for which two isoforms, short and long, have been described. Here we describe an additional isoform with a different RNA recognition motif, which is differentially expressed in the brain and ovary. We show that all CPEB1 isoforms are found associated with two previously described cytoplasmic structures, stress granules and dcp1 bodies. This association requires the RNA binding ability of the protein, whereas the Aurora A phosphorylation site is dispensable. Interestingly, the rck/p54 DEAD box protein, which is known as a CPEB partner in Xenopus and clam, and as a component of dcp1 bodies in mammals, is also present in stress granules. Both stress granules and dcp1 bodies are involved in mRNA storage and/or degradation, although so far no link has been made between the two, in terms of neither morphology nor protein content. Here we show that transient CPEB1 expression induces the assembly of stress granules, which in turn recruit dcp1 bodies. This dynamic connection between the two structures sheds new light on the compartmentalization of mRNA metabolism in the cytoplasm.

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“…This plasmid has a Rous sarcoma virus promoter-driven IL-3 cDNA and a hygromycin Bresistance gene (from pY3 plasmid) in the pBMGNeo backbone. [26][27][28][29] Bicistronic mammalian expression vectors…”

Section: Plasmid Constructionmentioning

confidence: 99%

Development of a modified selective amplifier gene for hematopoietic stem cell gene therapy

et al. 1999

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Hygromycin B phosphotransferase as a selectable marker for DNA transfer experiments with higher eucaryotic cells.

Cited by 254 publications

References 12 publications

Targeted inactivation of the insulin receptor gene in mouse 3T3-L1 fibroblasts via homologous recombination.

Targeted inactivation of the insulin receptor gene in mouse 3T3-L1 fibroblasts via homologous recombination.

The translational regulator CPEB1 provides a link between dcp1 bodies and stress granules

Development of a modified selective amplifier gene for hematopoietic stem cell gene therapy

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