{"title":"Hsp70A-RBCS2 融合启动子调控PHB合成酶基因在莱茵衣藻中的表达","authors":"王潮岗, 胡章立","doi":"10.1360/ZC2009-39-8-775","DOIUrl":null,"url":null,"abstract":"利用 Hsp70A-RBCS2 融合启动子构建了新型的莱茵衣藻表达载体, 并获得了聚-β-羟基丁酸(PHB)合成酶基因(phbC)的衣藻表达载体p105C124和pH105C124. 通过“珠磨法”分别将上述两个衣藻表达载体导入细胞壁缺陷的莱茵衣藻CC-849 ( Chlamydomonas reinhardtii CC-849)中, 得到了具有Zeomycin抗性的转基因藻株. Hsp70A-RBCS2启动子介导的外源基因遗传转化效率明显高于RBCS2启动子, Southern杂交结果显示, phbC基因以低拷贝数整合进莱茵衣藻的基因组DNA中. 在光照下, Hsp70A-RBCS2融合启动子能够有效调控phbC基因在莱茵衣藻中的转录和翻译, 得到的蛋白产物具有PHB合成酶活性, 40℃热激诱导可使PHB合成酶的酶活性提高到1.8倍. 因此, Hsp70A-RBCS2 融合启动子可使phbC基因在莱茵衣藻中实现可诱导的表达, 该研究对利用衣藻合成PHB具有重要的学术意义, 进一步的研究将通过“共转化”法获得二价或三价的转基因藻, 最终实现在转基因藻中合成PHB.","PeriodicalId":49127,"journal":{"name":"Science in China. Series C, Life Sciences / Chinese Academy of Sciences","volume":"27 1","pages":"775-782"},"PeriodicalIF":0.0000,"publicationDate":"2009-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Hsp70A-RBCS2 融合启动子调控PHB合成酶基因在莱茵衣藻中的表达\",\"authors\":\"王潮岗, 胡章立\",\"doi\":\"10.1360/ZC2009-39-8-775\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"利用 Hsp70A-RBCS2 融合启动子构建了新型的莱茵衣藻表达载体, 并获得了聚-β-羟基丁酸(PHB)合成酶基因(phbC)的衣藻表达载体p105C124和pH105C124. 通过“珠磨法”分别将上述两个衣藻表达载体导入细胞壁缺陷的莱茵衣藻CC-849 ( Chlamydomonas reinhardtii CC-849)中, 得到了具有Zeomycin抗性的转基因藻株. Hsp70A-RBCS2启动子介导的外源基因遗传转化效率明显高于RBCS2启动子, Southern杂交结果显示, phbC基因以低拷贝数整合进莱茵衣藻的基因组DNA中. 在光照下, Hsp70A-RBCS2融合启动子能够有效调控phbC基因在莱茵衣藻中的转录和翻译, 得到的蛋白产物具有PHB合成酶活性, 40℃热激诱导可使PHB合成酶的酶活性提高到1.8倍. 因此, Hsp70A-RBCS2 融合启动子可使phbC基因在莱茵衣藻中实现可诱导的表达, 该研究对利用衣藻合成PHB具有重要的学术意义, 进一步的研究将通过“共转化”法获得二价或三价的转基因藻, 最终实现在转基因藻中合成PHB.\",\"PeriodicalId\":49127,\"journal\":{\"name\":\"Science in China. Series C, Life Sciences / Chinese Academy of Sciences\",\"volume\":\"27 1\",\"pages\":\"775-782\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2009-08-20\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Science in China. Series C, Life Sciences / Chinese Academy of Sciences\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1360/ZC2009-39-8-775\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Science in China. Series C, Life Sciences / Chinese Academy of Sciences","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1360/ZC2009-39-8-775","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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