Development of a Sindbis virus expression system that efficiently expresses green fluorescent protein in midguts of Aedes aegypti following per os infection
A double subgenomic Sindbis (dsSIN) virus, MRE/3′2 J/GFP, was constructed to efficiently express green fluorescent protein (GFP) in the midgut of Aedes aegypti following per os infection. The MRE/3′2 J/GFP RNA genome contained the nonstructural genes and cis‐acting sequences of the dsSIN virus, TE/3...
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| Veröffentlicht in: | Insect molecular biology 2000-02, Vol.9 (1), p.57-65 |
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| container_title | Insect molecular biology |
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| creator | Olson, K. E. Myles, K. M. Seabaugh, R. C. Higgs, S. Carlson, J. O. Beaty, B. J. |
| description | A double subgenomic Sindbis (dsSIN) virus, MRE/3′2 J/GFP, was constructed to efficiently express green fluorescent protein (GFP) in the midgut of Aedes aegypti following per os infection. The MRE/3′2 J/GFP RNA genome contained the nonstructural genes and cis‐acting sequences of the dsSIN virus, TE/3′2 J/GFP, but had the structural genes of MRE16 SIN virus. MRE/3′2 J/GFP virus, unlike TE/3′2 J/GFP virus, efficiently infected mosquitoes orally. At 1–2 days postinfection, GFP was observed as multiple foci of expression on the lumenal side of the midgut. At 10–12 days postinfection, thirteen of fifteen mosquitoes infected with MRE/3′2 J/GFP virus had high levels of GFP expression in the mosquito midgut. The MRE3′2 J dsSIN expression system should be an important tool for efficient gene expression in Ae. aegypti midguts. |
| doi_str_mv | 10.1046/j.1365-2583.2000.00162.x |
| format | Article |
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E. ; Myles, K. M. ; Seabaugh, R. C. ; Higgs, S. ; Carlson, J. O. ; Beaty, B. J.</creator><creatorcontrib>Olson, K. E. ; Myles, K. M. ; Seabaugh, R. C. ; Higgs, S. ; Carlson, J. O. ; Beaty, B. J.</creatorcontrib><description>A double subgenomic Sindbis (dsSIN) virus, MRE/3′2 J/GFP, was constructed to efficiently express green fluorescent protein (GFP) in the midgut of Aedes aegypti following per os infection. The MRE/3′2 J/GFP RNA genome contained the nonstructural genes and cis‐acting sequences of the dsSIN virus, TE/3′2 J/GFP, but had the structural genes of MRE16 SIN virus. MRE/3′2 J/GFP virus, unlike TE/3′2 J/GFP virus, efficiently infected mosquitoes orally. At 1–2 days postinfection, GFP was observed as multiple foci of expression on the lumenal side of the midgut. At 10–12 days postinfection, thirteen of fifteen mosquitoes infected with MRE/3′2 J/GFP virus had high levels of GFP expression in the mosquito midgut. 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E.</creatorcontrib><creatorcontrib>Myles, K. M.</creatorcontrib><creatorcontrib>Seabaugh, R. C.</creatorcontrib><creatorcontrib>Higgs, S.</creatorcontrib><creatorcontrib>Carlson, J. O.</creatorcontrib><creatorcontrib>Beaty, B. J.</creatorcontrib><title>Development of a Sindbis virus expression system that efficiently expresses green fluorescent protein in midguts of Aedes aegypti following per os infection</title><title>Insect molecular biology</title><addtitle>Insect Mol Biol</addtitle><description>A double subgenomic Sindbis (dsSIN) virus, MRE/3′2 J/GFP, was constructed to efficiently express green fluorescent protein (GFP) in the midgut of Aedes aegypti following per os infection. The MRE/3′2 J/GFP RNA genome contained the nonstructural genes and cis‐acting sequences of the dsSIN virus, TE/3′2 J/GFP, but had the structural genes of MRE16 SIN virus. MRE/3′2 J/GFP virus, unlike TE/3′2 J/GFP virus, efficiently infected mosquitoes orally. At 1–2 days postinfection, GFP was observed as multiple foci of expression on the lumenal side of the midgut. At 10–12 days postinfection, thirteen of fifteen mosquitoes infected with MRE/3′2 J/GFP virus had high levels of GFP expression in the mosquito midgut. The MRE3′2 J dsSIN expression system should be an important tool for efficient gene expression in Ae. aegypti midguts.</description><subject>Aedes aegypti</subject><subject>Animals</subject><subject>Cells, Cultured</subject><subject>Culex - genetics</subject><subject>Culex - virology</subject><subject>Culicidae</subject><subject>Digestive System</subject><subject>Freshwater</subject><subject>Gene Expression Regulation, Viral</subject><subject>Genes, Viral</subject><subject>green fluorescent protein</subject><subject>Green Fluorescent Proteins</subject><subject>Luminescent Proteins - genetics</subject><subject>Sindbis virus</subject><subject>Sindbis Virus - genetics</subject><subject>Viral Nonstructural Proteins - genetics</subject><subject>Viral Structural Proteins - genetics</subject><subject>Virology - methods</subject><issn>0962-1075</issn><issn>1365-2583</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc1u1DAUhS0EokPhFZBX7BJsZ-wkEptOodNWw48EiKXlONeDhyRO7aSdvEsfFoeUiiWSJfvY37n3WgchTElKyVq8PaQ0EzxhvMhSRghJCaGCpccnaPX48BStSClYQknOT9CLEA4RLEpRPkcnlIickZyt0P17uIXG9S10A3YGK_zVdnVlA761fgwYjr2HEKzrcJjCAC0efqoBgzFW2-hppr8IBLz3AB02zejihZ4r9t4NYDscV2vr_TiEuckZ1JFWsJ_6wWLjmsbd2W6Pe_DYhQgb0ENs-RI9M6oJ8OphP0XfLz58O79Mdp-3V-dnu0SvCYs_5MaUSmujmeCk4DVwpo1idcVzUZSEZGW1pkLNSlGTRb2Oikay1LrS2Sl6s9SN496MEAbZ2jh_06gO3BgkzTnNWJ5HsFhA7V0IHozsvW2VnyQlck5GHuQcgJwDkHMy8k8y8hitrx96jFUL9T_GJYoIvFuAO9vA9N-F5dXHTTxEe7LYbUzp-GhX_pcUeZZz-ePTVm6u6fbiy_VG7rLfikWvsA</recordid><startdate>200002</startdate><enddate>200002</enddate><creator>Olson, K. 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E.</au><au>Myles, K. M.</au><au>Seabaugh, R. C.</au><au>Higgs, S.</au><au>Carlson, J. O.</au><au>Beaty, B. J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of a Sindbis virus expression system that efficiently expresses green fluorescent protein in midguts of Aedes aegypti following per os infection</atitle><jtitle>Insect molecular biology</jtitle><addtitle>Insect Mol Biol</addtitle><date>2000-02</date><risdate>2000</risdate><volume>9</volume><issue>1</issue><spage>57</spage><epage>65</epage><pages>57-65</pages><issn>0962-1075</issn><eissn>1365-2583</eissn><abstract>A double subgenomic Sindbis (dsSIN) virus, MRE/3′2 J/GFP, was constructed to efficiently express green fluorescent protein (GFP) in the midgut of Aedes aegypti following per os infection. The MRE/3′2 J/GFP RNA genome contained the nonstructural genes and cis‐acting sequences of the dsSIN virus, TE/3′2 J/GFP, but had the structural genes of MRE16 SIN virus. 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| source | MEDLINE; Wiley Online Library Journals Frontfile Complete |
| subjects | Aedes aegypti Animals Cells, Cultured Culex - genetics Culex - virology Culicidae Digestive System Freshwater Gene Expression Regulation, Viral Genes, Viral green fluorescent protein Green Fluorescent Proteins Luminescent Proteins - genetics Sindbis virus Sindbis Virus - genetics Viral Nonstructural Proteins - genetics Viral Structural Proteins - genetics Virology - methods |
| title | Development of a Sindbis virus expression system that efficiently expresses green fluorescent protein in midguts of Aedes aegypti following per os infection |
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