Promoter of shrimp translationally controlled tumor protein (TCTP) gene has higher activity than cytomegavirus (CMV) promoter in driving foreign genes to express in primary shrimp cells
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Abstract
Active promoters are urgently needed for shrimp cells which are hard to be immortalized. Translationally controlled tumor protein (TCTP) is a widely and abundantly expressed, growth-related protein. In this study we successfully isolated the promoter (Ptctp) and ORF (= open reading frame) of the TCTP gene from Litopenaeus vannamei and analysed the promoter activity of Ptctp and the growth-promoting effects of over-expressed TCTP protein in the primary Oka organ (lymphoid tissue) cells. It was found, that cytomegavirus (CMV) promoter, highly active in mammalian and fish cells, had a driving activity too low to be detected in shrimp cells. However, shrimp Ptctp had a higher driving activity than Pcmv in the shrimp cells and an obvious fluorescent signal was observed in the shrimp cells transfected by a Pcmv-Ptctp-driven plasmid. However, no obvious growth-promoting effects were observed in the eGFP/TCTP [eGFP = enhanced green fluorescent protein] or TCTP-transfected shrimp cells possibly due to the relatively low expression efficiency.
Résumé
Des promoteurs actifs sont nécessaires pour les cellules de crevettes qui sont difficiles à immortaliser. La protéine TCTP (Translationally controlled tumor protein) est une protéine largement et abondamment représentée, relié à la croissance. Dans cette étude nous avons avec succès isolé le promoteur (Ptctp) et ORF (= cadre de lecture ouverte) du gène TCTP de Litopenaeus vannamei et analysé l’activité du promoteur de Ptctp et les effets promoteurs sur la croissance de la protéine TCTP sur-exprimée dans les cellules du tissu lymphoïde. Il a été trouvé que le promoteur du cytomégalovirus (CMV), fortement actif chez les cellules de mammifères et de poissons, a un niveau d’activité trop faible pour être détecté dans les cellules de crevette. Cependant le Ptctp crevette a présenté un plus fort niveau d’activité que Pcmv dans les cellules de crevettes et un signal fluorescent évident a été observé dans les cellules de crevette transfectées par un plasmide Pcmv-Ptctp. Cependant, aucun effet de promotion de croissance n’a été observé sur eGFP/TCTP ou dans les cellules de crevette TCTP-transfectées, sans doute à cause d’une efficacité de l’expression relativement faible.
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Promoter of shrimp translationally controlled tumor protein (TCTP) gene has higher activity than cytomegavirus (CMV) promoter in driving foreign genes to express in primary shrimp cells
In: CrustaceanaSearch for other papers by Jing Wang in
Current site
Google Scholar
PubMed
Search for other papers by Yuqi Lin in
Current site
Google Scholar
PubMed
Search for other papers by Xin Song in
Current site
Google Scholar
PubMed
Search for other papers by Longjun Pu in
Current site
Google Scholar
PubMed
Search for other papers by Guangdong Ji in
Current site
Google Scholar
PubMed
Search for other papers by Guorong Li in
Current site
Google Scholar
PubMed
Institute of Evolution and Marine Biodiversity, Ocean University of China, Qingdao 266003, P.R. China
Search for other papers by Huarong Guo in
Current site
Google Scholar
PubMed
Abstract
Active promoters are urgently needed for shrimp cells which are hard to be immortalized. Translationally controlled tumor protein (TCTP) is a widely and abundantly expressed, growth-related protein. In this study we successfully isolated the promoter (Ptctp) and ORF (= open reading frame) of the TCTP gene from Litopenaeus vannamei and analysed the promoter activity of Ptctp and the growth-promoting effects of over-expressed TCTP protein in the primary Oka organ (lymphoid tissue) cells. It was found, that cytomegavirus (CMV) promoter, highly active in mammalian and fish cells, had a driving activity too low to be detected in shrimp cells. However, shrimp Ptctp had a higher driving activity than Pcmv in the shrimp cells and an obvious fluorescent signal was observed in the shrimp cells transfected by a Pcmv-Ptctp-driven plasmid. However, no obvious growth-promoting effects were observed in the eGFP/TCTP [eGFP = enhanced green fluorescent protein] or TCTP-transfected shrimp cells possibly due to the relatively low expression efficiency.
Résumé
Des promoteurs actifs sont nécessaires pour les cellules de crevettes qui sont difficiles à immortaliser. La protéine TCTP (Translationally controlled tumor protein) est une protéine largement et abondamment représentée, relié à la croissance. Dans cette étude nous avons avec succès isolé le promoteur (Ptctp) et ORF (= cadre de lecture ouverte) du gène TCTP de Litopenaeus vannamei et analysé l’activité du promoteur de Ptctp et les effets promoteurs sur la croissance de la protéine TCTP sur-exprimée dans les cellules du tissu lymphoïde. Il a été trouvé que le promoteur du cytomégalovirus (CMV), fortement actif chez les cellules de mammifères et de poissons, a un niveau d’activité trop faible pour être détecté dans les cellules de crevette. Cependant le Ptctp crevette a présenté un plus fort niveau d’activité que Pcmv dans les cellules de crevettes et un signal fluorescent évident a été observé dans les cellules de crevette transfectées par un plasmide Pcmv-Ptctp. Cependant, aucun effet de promotion de croissance n’a été observé sur eGFP/TCTP ou dans les cellules de crevette TCTP-transfectées, sans doute à cause d’une efficacité de l’expression relativement faible.
| All Time | Past 365 days | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 709 | 247 | 22 |
| Full Text Views | 26 | 0 | 0 |
| PDF Views & Downloads | 32 | 1 | 0 |
