Transient gene expression in electroporated Picea glauca protoplasts
The reporter gene for chloramphenicol acetyltransferase (CAT) was introduced into white spruce (Picea glauca (Moench) Voss.) protoplasts by electroporation. CAT transient gene expression was increased by increasing the concentration of pCaMVCN plasmid and was affected by the level of the applied vol...
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| Veröffentlicht in: | Plant cell reports 1988-12, Vol.7 (7), p.481-484 |
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| container_title | Plant cell reports |
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| creator | Bekkaoui, F Pilon, M Laine, E Raju, D.S.S Crosby, W.L Dunstan, D.I |
| description | The reporter gene for chloramphenicol acetyltransferase (CAT) was introduced into white spruce (Picea glauca (Moench) Voss.) protoplasts by electroporation. CAT transient gene expression was increased by increasing the concentration of pCaMVCN plasmid and was affected by the level of the applied voltage. Highest CAT activities were obtained after electroporation with a pulse of 350V.cm(-1) having an exponential decay constant of approximately 105ms. Linearized plasmid constructs gave much higher levels of CAT activity than circular plasmid. Attempts to use the Escherichia coli β-glucuronidase gene (β-GUS) as a marker gene revealed very high levels of β-GUS-like activity in electroporated protoplasts. This activity was mainly due to a small molecule and may mask successful transformation since β-GUS-like activity increased when plasmid DNA was present during electroporation. |
| doi_str_mv | 10.1007/BF00272736 |
| format | Article |
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CAT transient gene expression was increased by increasing the concentration of pCaMVCN plasmid and was affected by the level of the applied voltage. Highest CAT activities were obtained after electroporation with a pulse of 350V.cm(-1) having an exponential decay constant of approximately 105ms. Linearized plasmid constructs gave much higher levels of CAT activity than circular plasmid. Attempts to use the Escherichia coli β-glucuronidase gene (β-GUS) as a marker gene revealed very high levels of β-GUS-like activity in electroporated protoplasts. This activity was mainly due to a small molecule and may mask successful transformation since β-GUS-like activity increased when plasmid DNA was present during electroporation.</description><identifier>ISSN: 0721-7714</identifier><identifier>EISSN: 1432-203X</identifier><identifier>DOI: 10.1007/BF00272736</identifier><identifier>PMID: 24240396</identifier><identifier>CODEN: PCRPD8</identifier><language>eng</language><publisher>Berlin: Springer</publisher><subject>Biological and medical sciences ; Biotechnology ; chloramphenicol ; enzyme activity ; Fundamental and applied biological sciences. Psychology ; Gene expression ; Genetic engineering ; Genetic technics ; genetic transformation ; Methods. Procedures. Technologies ; Miscellaneous ; Molecular and cellular biology ; Molecular genetics ; Picea glauca ; plasmids ; protoplast fusion ; Synthetic digonucleotides and genes. 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CAT transient gene expression was increased by increasing the concentration of pCaMVCN plasmid and was affected by the level of the applied voltage. Highest CAT activities were obtained after electroporation with a pulse of 350V.cm(-1) having an exponential decay constant of approximately 105ms. Linearized plasmid constructs gave much higher levels of CAT activity than circular plasmid. Attempts to use the Escherichia coli β-glucuronidase gene (β-GUS) as a marker gene revealed very high levels of β-GUS-like activity in electroporated protoplasts. This activity was mainly due to a small molecule and may mask successful transformation since β-GUS-like activity increased when plasmid DNA was present during electroporation.</description><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>chloramphenicol</subject><subject>enzyme activity</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene expression</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>genetic transformation</subject><subject>Methods. Procedures. Technologies</subject><subject>Miscellaneous</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Picea glauca</subject><subject>plasmids</subject><subject>protoplast fusion</subject><subject>Synthetic digonucleotides and genes. 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Psychology</topic><topic>Gene expression</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>genetic transformation</topic><topic>Methods. Procedures. Technologies</topic><topic>Miscellaneous</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Picea glauca</topic><topic>plasmids</topic><topic>protoplast fusion</topic><topic>Synthetic digonucleotides and genes. Sequencing</topic><topic>transferases</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bekkaoui, F</creatorcontrib><creatorcontrib>Pilon, M</creatorcontrib><creatorcontrib>Laine, E</creatorcontrib><creatorcontrib>Raju, D.S.S</creatorcontrib><creatorcontrib>Crosby, W.L</creatorcontrib><creatorcontrib>Dunstan, D.I</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Plant cell reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bekkaoui, F</au><au>Pilon, M</au><au>Laine, E</au><au>Raju, D.S.S</au><au>Crosby, W.L</au><au>Dunstan, D.I</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Transient gene expression in electroporated Picea glauca protoplasts</atitle><jtitle>Plant cell reports</jtitle><addtitle>Plant Cell Rep</addtitle><date>1988-12-01</date><risdate>1988</risdate><volume>7</volume><issue>7</issue><spage>481</spage><epage>484</epage><pages>481-484</pages><issn>0721-7714</issn><eissn>1432-203X</eissn><coden>PCRPD8</coden><abstract>The reporter gene for chloramphenicol acetyltransferase (CAT) was introduced into white spruce (Picea glauca (Moench) Voss.) protoplasts by electroporation. CAT transient gene expression was increased by increasing the concentration of pCaMVCN plasmid and was affected by the level of the applied voltage. Highest CAT activities were obtained after electroporation with a pulse of 350V.cm(-1) having an exponential decay constant of approximately 105ms. Linearized plasmid constructs gave much higher levels of CAT activity than circular plasmid. Attempts to use the Escherichia coli β-glucuronidase gene (β-GUS) as a marker gene revealed very high levels of β-GUS-like activity in electroporated protoplasts. This activity was mainly due to a small molecule and may mask successful transformation since β-GUS-like activity increased when plasmid DNA was present during electroporation.</abstract><cop>Berlin</cop><pub>Springer</pub><pmid>24240396</pmid><doi>10.1007/BF00272736</doi><tpages>4</tpages></addata></record> |
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| ispartof | Plant cell reports, 1988-12, Vol.7 (7), p.481-484 |
| issn | 0721-7714 1432-203X |
| language | eng |
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| source | SpringerLink Journals - AutoHoldings |
| subjects | Biological and medical sciences Biotechnology chloramphenicol enzyme activity Fundamental and applied biological sciences. Psychology Gene expression Genetic engineering Genetic technics genetic transformation Methods. Procedures. Technologies Miscellaneous Molecular and cellular biology Molecular genetics Picea glauca plasmids protoplast fusion Synthetic digonucleotides and genes. Sequencing transferases |
| title | Transient gene expression in electroporated Picea glauca protoplasts |
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