本文關鍵詞：乙肝病毒表面抗原基因在人參細胞中的高效表達　出處：《吉林大學》2007年博士論文　論文類型：學位論文

  更多相關文章： 翻譯增強子 omega序列 乙肝病毒表面抗原 人參愈傷組織

【摘要】： 我們在前期工作中建立了人參愈傷組織細胞表達乙肝病毒表面抗原(HBsAg)的植物表達系統(tǒng),并就提高表達進行了培養(yǎng)條件的摸索。為了進一步提高HBsAg的表達,我們從表達載體的改造上做了一些探索,對調控HBsAg基因表達的啟動子以及翻譯增強子進行了改造,構建了十二種不同的植物細胞表達載體pBI/EP、pBI/2EP、pBI/3EP、pBI/4EP、pBI/5EP、pBI/6EP、pBI/7EP、pBI/8EP、pBIBSa(pBI35S)、pBI/2EPomega、pBI/35Somega和pBI/35SRomega,用這些載體分別轉化農桿菌LBA4404,再與人參愈傷組織細胞共培養(yǎng),經G418篩選獲得新生抗性細胞,從而將HBsAg基因以及調控基因整合到人參細胞基因組中,經過連續(xù)繼代培養(yǎng)選育出一株高產細胞系PBIo。 PBIo是由質粒pBI/35Somega轉化的人參細胞選育而來,攜帶花椰菜花葉病毒CaMV 35S啟動子以及與之串聯(lián)的煙草花葉病毒TMV(U1株)翻譯增強子omega序列。通過對表達載體的改造使HBsAg基因表達有顯著提高。在實驗中還發(fā)現,omega反義RNA序列也具有較強的提高翻譯效率的功能。
[Abstract]:A plant expression system for the expression of hepatitis B virus surface antigen (HBsAg) in callus cells of Panax ginseng was established in our earlier work. In order to further improve the expression of HBsAg, we made some exploration on the transformation of expression vector. The promoter and the translation enhancer regulating the expression of HBsAg gene were modified and 12 different plant cell expression vectors pBI / EPP / 2 EPBI / 3 EP were constructed. PBI / 4EPB / 5EPBI / 6EPBI / 7EPBI / 8EPBI / 8EPBI35Sv PBI / 2EPomega; PBI / PBI / 5EPBI / 5EPBI / 6EPB / 7EPBI / 8EPBI / 8EPBI35Sv / PBI / 2EPomega. PBI/35Somega and pBI / 35 SRomega were transformed into Agrobacterium tumefaciens LBA4404 respectively and co-cultured with ginseng callus cells. New resistant cells were obtained by G418 screening, and then the HBsAg gene and regulatory gene were integrated into the genome of ginseng cells, and a high-yielding cell line PBIoO was selected after successive subculture. PBIo was selected from ginseng cells transformed by plasmid pBI/35Somega. Carrying CaMV 35s promoter of cauliflower mosaic virus and TMV(U1 strain of tobacco mosaic virus (TMV). Translation enhancer omega sequence. Through the modification of expression vector, the expression of HBsAg gene was significantly improved. It was also found in the experiment. Omega antisense RNA sequences can also improve translation efficiency.
【學位授予單位】：吉林大學
【學位級別】：博士
【學位授予年份】：2007
【分類號】：R392;Q943.2

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相關博士學位論文 前1條

1 高正倫;乙肝病毒表面抗原基因在人參細胞中的高效表達[D];吉林大學;2007年

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