Publications related to Molecular cloning

Production of recombinant protein therapeutics in cultivated mammalian cells

Cultivated mammalian cells have become the dominant system for the production of recombinant proteins for clinical applications because of their capacity for proper protein folding, assembly and post-translational modification. Thus, the quality and effica ...

2004

Transfection of partially purified plasmid DNA for high level transient protein expression in HEK293-EBNA cells

Florian Maria Wurm, Martin Jordan, Jason Wright

One of the major constraints to performing large-scale transfections of cultured mammalian cells for the transient expression of recombinant proteins is the production of large quantities of purified plasmid DNA. In this report partially purified plasmid D ...

2003

Correlation between cell cycle and calcium phosphate transient transfection of CHO cells

Frédéric Grosjean

Calcium phosphate transfection is a widely used method to produce recombinant proteins in mammalian cells. However, the mechanisms involved in plasmid DNA transfer to the nucleus of the transfected cells remain poorly understood and result in great variati ...

EPFL2003

Cloning of Baeyer-Villiger monooxygenases from Comamonas, Xanthobacter and Thodococcus using polymerase chain reaction with highly degenerate primers

To clone novel type 1 Baeyer-Villiger monooxygenase (BVMO) genes, we isolated or collected 25 bacterial strains able to grow on alicyclic compounds. Twelve of the bacterial strains yielded polymerase chain reaction (PCR) fragments with highly degenerate pr ...

2003

Structural Arrest in Dense Star-Polymer Solutions

Giuseppe Foffi, Francesco Sciortino

The dynamics of star polymers is investigated via extensive molecular and Brownian dynamics simulations for a large range of functionality f and packing fraction

\eta

. The calculated isodiffusivity curves display both minima and maxima as a function of $ ...

2003

Transfection using plasmid preparations

Florian Maria Wurm, Jason Wright

There is provided a method for plasmid DNA extraction to reproducibly yield relatively crude preparation which is suitable for transfection into mammalian cells. The inventors have determined that the high level of purification previously thought to be nec ...

2002

Fluorescent proteins in animal cells for process development: optimization of sodium butyrate treatment as an example

Florian Maria Wurm, Martin Jordan, Lisa Hunt

Fluorescent proteins expressed in mammalian cells can be quantified quickly and noninvasively with a standard fluorescence plate reader. We have previously exploited this quality in cell growth assessment (Hunt et al., 1999b). In this work, different CHO c ...

2002

Balancing GFP reporter plasmid quantity in large-scale transient transfections for recombinant anti-human Rhesus-D IgG1 synthesis

Horst Vogel, Florian Maria Wurm, Horst Pick

Using transient expression, high amounts (>20 mg/mL) of secreted anti-human Rhesus-D IgG1 were produced in a suspension-adapted HEK293 EBNA cell line (Meissner et al., Biotechnol Bioeng 75: 197-203, 2001). Time of harvest was 3 days after transfection. For ...

2002

Development of stable cell lines for production or regulated expression using matrix attachment regions

Florian Maria Wurm

One of the major hurdles of isolating stable, inducible or constitutive high-level producer cell lines is the time-consuming selection procedure. Given the variation in the expression levels of the same construct in individual clones, hundreds of clones mu ...

2001

Molecular cloning, genetic mapping, and expression analysis of four zebrafish c/ebp genes

Andrew Charles Oates

The CCAAT/enhancer binding protein family (C/EBP) are transcription factors that play integral roles in the development and function of many organ systems, including hematopoietic cells, adipose tissues, and liver. We have identified and characterized puta ...

2001

Molecular cloning

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Production of recombinant protein therapeutics in cultivated mammalian cells

Transfection of partially purified plasmid DNA for high level transient protein expression in HEK293-EBNA cells

Correlation between cell cycle and calcium phosphate transient transfection of CHO cells

Cloning of Baeyer-Villiger monooxygenases from Comamonas, Xanthobacter and Thodococcus using polymerase chain reaction with highly degenerate primers

Structural Arrest in Dense Star-Polymer Solutions

Transfection using plasmid preparations

Fluorescent proteins in animal cells for process development: optimization of sodium butyrate treatment as an example

Balancing GFP reporter plasmid quantity in large-scale transient transfections for recombinant anti-human Rhesus-D IgG1 synthesis

Development of stable cell lines for production or regulated expression using matrix attachment regions

Molecular cloning, genetic mapping, and expression analysis of four zebrafish c/ebp genes

Development of stable cell lines for production or regulated expression using matrix attachment regions

Fluorescent proteins in animal cells for process development: optimization of sodium butyrate treatment as an example

Production of recombinant protein therapeutics in cultivated mammalian cells

Transfection using plasmid preparations

Balancing GFP reporter plasmid quantity in large-scale transient transfections for recombinant anti-human Rhesus-D IgG1 synthesis

Molecular cloning, genetic mapping, and expression analysis of four zebrafish c/ebp genes

Transfection of partially purified plasmid DNA for high level transient protein expression in HEK293-EBNA cells

Cloning of Baeyer-Villiger monooxygenases from Comamonas, Xanthobacter and Thodococcus using polymerase chain reaction with highly degenerate primers

Correlation between cell cycle and calcium phosphate transient transfection of CHO cells

Structural Arrest in Dense Star-Polymer Solutions