{"title":"[刚地弓形虫重组质粒pcDNA3/GRA1的构建与测序]。","authors":"Li-ting Cai, Heng-ping Shu, Li-ping Jiang","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To construct a mammalian expression plasmid pcDNA3/GRA1 to express dense granules antigen-1 (GRA1) of Toxoplasma gondii, and to lay a foundation for further studying the protective immunity of pcDNA3/GRA1 as a DNA vaccine.</p><p><strong>Methods: </strong>The GRA1 opening reading frame (ORF) was amplified with two specific primers. The ORF and plasmid pcDNA3 were digested with EcoR I and Xho I respectively and the ORF was ligated into the pcDNA3 at polylinker. The recombinant vector pcDNA3/GRA1 was characterized by PCR, restriction enzyme digestion, and sequencing analysis.</p><p><strong>Results: </strong>The expected ORF, 573 bp long, was amplified by PCR, and inserted into plasmid pcDNA3. PCR, restriction enzyme digestion and sequencing analysis showed that pcDNA3/GRA1 contained GRA1 ORF with the right orientation.</p><p><strong>Conclusion: </strong>The mammalian expression vector pcDNA3/GRA1 is successfully constructed.</p>","PeriodicalId":13115,"journal":{"name":"Hunan yi ke da xue xue bao = Hunan yike daxue xuebao = Bulletin of Hunan Medical University","volume":"28 3","pages":"221-3"},"PeriodicalIF":0.0000,"publicationDate":"2003-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Construction and sequencing of recombinant plasmid pcDNA3/GRA1 from Toxoplasma gondii].\",\"authors\":\"Li-ting Cai, Heng-ping Shu, Li-ping Jiang\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>To construct a mammalian expression plasmid pcDNA3/GRA1 to express dense granules antigen-1 (GRA1) of Toxoplasma gondii, and to lay a foundation for further studying the protective immunity of pcDNA3/GRA1 as a DNA vaccine.</p><p><strong>Methods: </strong>The GRA1 opening reading frame (ORF) was amplified with two specific primers. The ORF and plasmid pcDNA3 were digested with EcoR I and Xho I respectively and the ORF was ligated into the pcDNA3 at polylinker. The recombinant vector pcDNA3/GRA1 was characterized by PCR, restriction enzyme digestion, and sequencing analysis.</p><p><strong>Results: </strong>The expected ORF, 573 bp long, was amplified by PCR, and inserted into plasmid pcDNA3. PCR, restriction enzyme digestion and sequencing analysis showed that pcDNA3/GRA1 contained GRA1 ORF with the right orientation.</p><p><strong>Conclusion: </strong>The mammalian expression vector pcDNA3/GRA1 is successfully constructed.</p>\",\"PeriodicalId\":13115,\"journal\":{\"name\":\"Hunan yi ke da xue xue bao = Hunan yike daxue xuebao = Bulletin of Hunan Medical University\",\"volume\":\"28 3\",\"pages\":\"221-3\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2003-06-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Hunan yi ke da xue xue bao = Hunan yike daxue xuebao = Bulletin of Hunan Medical University\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Hunan yi ke da xue xue bao = Hunan yike daxue xuebao = Bulletin of Hunan Medical University","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
[Construction and sequencing of recombinant plasmid pcDNA3/GRA1 from Toxoplasma gondii].
Objective: To construct a mammalian expression plasmid pcDNA3/GRA1 to express dense granules antigen-1 (GRA1) of Toxoplasma gondii, and to lay a foundation for further studying the protective immunity of pcDNA3/GRA1 as a DNA vaccine.
Methods: The GRA1 opening reading frame (ORF) was amplified with two specific primers. The ORF and plasmid pcDNA3 were digested with EcoR I and Xho I respectively and the ORF was ligated into the pcDNA3 at polylinker. The recombinant vector pcDNA3/GRA1 was characterized by PCR, restriction enzyme digestion, and sequencing analysis.
Results: The expected ORF, 573 bp long, was amplified by PCR, and inserted into plasmid pcDNA3. PCR, restriction enzyme digestion and sequencing analysis showed that pcDNA3/GRA1 contained GRA1 ORF with the right orientation.
Conclusion: The mammalian expression vector pcDNA3/GRA1 is successfully constructed.
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