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Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Larg...
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| Zeitschriftentitel: | Gene Regulation and Systems Biology |
|---|---|
| Personen und Körperschaften: | , , , |
| In: | Gene Regulation and Systems Biology, 1, 2007, S. 117762500700100 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
SAGE Publications
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| Schlagwörter: |
| author_facet |
Tröße, Christiane Ravneberg, Hanne Stern, Beate Pryme, Ian F. Tröße, Christiane Ravneberg, Hanne Stern, Beate Pryme, Ian F. |
|---|---|
| author |
Tröße, Christiane Ravneberg, Hanne Stern, Beate Pryme, Ian F. |
| spellingShingle |
Tröße, Christiane Ravneberg, Hanne Stern, Beate Pryme, Ian F. Gene Regulation and Systems Biology Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected Computer Science Applications Genetics Molecular Biology Ecology, Evolution, Behavior and Systematics |
| author_sort |
tröße, christiane |
| spelling |
Tröße, Christiane Ravneberg, Hanne Stern, Beate Pryme, Ian F. 1177-6250 1177-6250 SAGE Publications Computer Science Applications Genetics Molecular Biology Ecology, Evolution, Behavior and Systematics http://dx.doi.org/10.1177/117762500700100021 <jats:p>The signal peptide of the luciferase secreted by the marine copepod Gaussia princeps has been shown to promote high-level protein synthesis/secretion of recombinant proteins, being far superior to mammalian counterparts. The main aim of the present study was to investigate the effects of seven selected signal peptides derived from oikosins, house proteins of the marine organism Oikopleura dioica, on synthesis/secretion of recombinant proteins. Vector constructs were made in which the coding regions of two naturally secreted proteins, Gaussia luciferase and human endostatin (hEndostatin), were “seamlessly” fused to the signal peptide coding sequences of interest. CHO cells were transfected with the plasmids and populations of stably transfected cells established. The amounts of reporter proteins in cell extract and medium samples were determined and the results compared to those obtained from cells stably transfected with a reference vector construct. In addition, the amounts of luciferase or hEndostatin encoding mRNAs in the cells were determined and related to the protein levels obtained. The levels of reporter protein produced varied greatly among the seven oikosin signal peptides tested. Whereas the oikosin 1 signal peptide resulted in about 40% production of Gaussia luciferase compared to the reference construct, oikosins 2–7 were extremely ineffective (<1%). mRNA levels were not dramatically affected such that inadequate availability of transcript for translation was not the underlying reason for the observations. The oikosin 1 signal peptide was also the most effective regarding synthesis/secretion of hEndostatin. No secreted product was observed using the oikosin 3 signal peptide. Interestingly, the molecular weight of hEndostatin in cell extracts prepared from cells transfected with oikosin 2 and 3 constructs was higher than that using the oikosin 1 signal peptide. The overall findings indicate that the signal peptide affects the efficiency of protein synthesis and secretion through a mechanism operating at the post-transcriptional level. The results described here provide substantial support to our previous observations which suggested that the choice of the signal peptide is imperative when aiming to achieve optimal synthesis and secretion of a recombinant protein using transfected mammalian cells.</jats:p> Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in Mediating<i>Gaussia</i>luciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected Gene Regulation and Systems Biology |
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| title |
Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected |
| title_unstemmed |
Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected |
| title_full |
Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected |
| title_fullStr |
Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected |
| title_full_unstemmed |
Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected |
| title_short |
Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected |
| title_sort |
vectors encoding seven oikosin signal peptides transfected into cho cells differ greatly in mediating<i>gaussia</i>luciferase and human endostatin production although mrna levels are largely unaffected |
| topic |
Computer Science Applications Genetics Molecular Biology Ecology, Evolution, Behavior and Systematics |
| url |
http://dx.doi.org/10.1177/117762500700100021 |
| publishDate |
2007 |
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117762500700100 |
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<jats:p>The signal peptide of the luciferase secreted by the marine copepod Gaussia princeps has been shown to promote high-level protein synthesis/secretion of recombinant proteins, being far superior to mammalian counterparts. The main aim of the present study was to investigate the effects of seven selected signal peptides derived from oikosins, house proteins of the marine organism Oikopleura dioica, on synthesis/secretion of recombinant proteins. Vector constructs were made in which the coding regions of two naturally secreted proteins, Gaussia luciferase and human endostatin (hEndostatin), were “seamlessly” fused to the signal peptide coding sequences of interest. CHO cells were transfected with the plasmids and populations of stably transfected cells established. The amounts of reporter proteins in cell extract and medium samples were determined and the results compared to those obtained from cells stably transfected with a reference vector construct. In addition, the amounts of luciferase or hEndostatin encoding mRNAs in the cells were determined and related to the protein levels obtained. The levels of reporter protein produced varied greatly among the seven oikosin signal peptides tested. Whereas the oikosin 1 signal peptide resulted in about 40% production of Gaussia luciferase compared to the reference construct, oikosins 2–7 were extremely ineffective (<1%). mRNA levels were not dramatically affected such that inadequate availability of transcript for translation was not the underlying reason for the observations. The oikosin 1 signal peptide was also the most effective regarding synthesis/secretion of hEndostatin. No secreted product was observed using the oikosin 3 signal peptide. Interestingly, the molecular weight of hEndostatin in cell extracts prepared from cells transfected with oikosin 2 and 3 constructs was higher than that using the oikosin 1 signal peptide. The overall findings indicate that the signal peptide affects the efficiency of protein synthesis and secretion through a mechanism operating at the post-transcriptional level. The results described here provide substantial support to our previous observations which suggested that the choice of the signal peptide is imperative when aiming to achieve optimal synthesis and secretion of a recombinant protein using transfected mammalian cells.</jats:p> |
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| author | Tröße, Christiane, Ravneberg, Hanne, Stern, Beate, Pryme, Ian F. |
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| description | <jats:p>The signal peptide of the luciferase secreted by the marine copepod Gaussia princeps has been shown to promote high-level protein synthesis/secretion of recombinant proteins, being far superior to mammalian counterparts. The main aim of the present study was to investigate the effects of seven selected signal peptides derived from oikosins, house proteins of the marine organism Oikopleura dioica, on synthesis/secretion of recombinant proteins. Vector constructs were made in which the coding regions of two naturally secreted proteins, Gaussia luciferase and human endostatin (hEndostatin), were “seamlessly” fused to the signal peptide coding sequences of interest. CHO cells were transfected with the plasmids and populations of stably transfected cells established. The amounts of reporter proteins in cell extract and medium samples were determined and the results compared to those obtained from cells stably transfected with a reference vector construct. In addition, the amounts of luciferase or hEndostatin encoding mRNAs in the cells were determined and related to the protein levels obtained. The levels of reporter protein produced varied greatly among the seven oikosin signal peptides tested. Whereas the oikosin 1 signal peptide resulted in about 40% production of Gaussia luciferase compared to the reference construct, oikosins 2–7 were extremely ineffective (<1%). mRNA levels were not dramatically affected such that inadequate availability of transcript for translation was not the underlying reason for the observations. The oikosin 1 signal peptide was also the most effective regarding synthesis/secretion of hEndostatin. No secreted product was observed using the oikosin 3 signal peptide. Interestingly, the molecular weight of hEndostatin in cell extracts prepared from cells transfected with oikosin 2 and 3 constructs was higher than that using the oikosin 1 signal peptide. The overall findings indicate that the signal peptide affects the efficiency of protein synthesis and secretion through a mechanism operating at the post-transcriptional level. The results described here provide substantial support to our previous observations which suggested that the choice of the signal peptide is imperative when aiming to achieve optimal synthesis and secretion of a recombinant protein using transfected mammalian cells.</jats:p> |
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| institution | DE-Bn3, DE-Brt1, DE-D161, DE-Zwi2, DE-Gla1, DE-Zi4, DE-15, DE-Pl11, DE-Rs1, DE-105, DE-14, DE-Ch1, DE-L229, DE-D275 |
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| spelling | Tröße, Christiane Ravneberg, Hanne Stern, Beate Pryme, Ian F. 1177-6250 1177-6250 SAGE Publications Computer Science Applications Genetics Molecular Biology Ecology, Evolution, Behavior and Systematics http://dx.doi.org/10.1177/117762500700100021 <jats:p>The signal peptide of the luciferase secreted by the marine copepod Gaussia princeps has been shown to promote high-level protein synthesis/secretion of recombinant proteins, being far superior to mammalian counterparts. The main aim of the present study was to investigate the effects of seven selected signal peptides derived from oikosins, house proteins of the marine organism Oikopleura dioica, on synthesis/secretion of recombinant proteins. Vector constructs were made in which the coding regions of two naturally secreted proteins, Gaussia luciferase and human endostatin (hEndostatin), were “seamlessly” fused to the signal peptide coding sequences of interest. CHO cells were transfected with the plasmids and populations of stably transfected cells established. The amounts of reporter proteins in cell extract and medium samples were determined and the results compared to those obtained from cells stably transfected with a reference vector construct. In addition, the amounts of luciferase or hEndostatin encoding mRNAs in the cells were determined and related to the protein levels obtained. The levels of reporter protein produced varied greatly among the seven oikosin signal peptides tested. Whereas the oikosin 1 signal peptide resulted in about 40% production of Gaussia luciferase compared to the reference construct, oikosins 2–7 were extremely ineffective (<1%). mRNA levels were not dramatically affected such that inadequate availability of transcript for translation was not the underlying reason for the observations. The oikosin 1 signal peptide was also the most effective regarding synthesis/secretion of hEndostatin. No secreted product was observed using the oikosin 3 signal peptide. Interestingly, the molecular weight of hEndostatin in cell extracts prepared from cells transfected with oikosin 2 and 3 constructs was higher than that using the oikosin 1 signal peptide. The overall findings indicate that the signal peptide affects the efficiency of protein synthesis and secretion through a mechanism operating at the post-transcriptional level. The results described here provide substantial support to our previous observations which suggested that the choice of the signal peptide is imperative when aiming to achieve optimal synthesis and secretion of a recombinant protein using transfected mammalian cells.</jats:p> Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in Mediating<i>Gaussia</i>luciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected Gene Regulation and Systems Biology |
| spellingShingle | Tröße, Christiane, Ravneberg, Hanne, Stern, Beate, Pryme, Ian F., Gene Regulation and Systems Biology, Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected, Computer Science Applications, Genetics, Molecular Biology, Ecology, Evolution, Behavior and Systematics |
| title | Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected |
| title_full | Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected |
| title_fullStr | Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected |
| title_full_unstemmed | Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected |
| title_short | Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected |
| title_sort | vectors encoding seven oikosin signal peptides transfected into cho cells differ greatly in mediating<i>gaussia</i>luciferase and human endostatin production although mrna levels are largely unaffected |
| title_unstemmed | Vectors Encoding Seven Oikosin Signal Peptides Transfected into CHO Cells Differ Greatly in MediatingGaussialuciferase and Human Endostatin Production although mRNA Levels are Largely Unaffected |
| topic | Computer Science Applications, Genetics, Molecular Biology, Ecology, Evolution, Behavior and Systematics |
| url | http://dx.doi.org/10.1177/117762500700100021 |