本文關(guān)鍵詞：乙肝病毒表面抗原基因在人參細(xì)胞中的高效表達(dá)　出處：《吉林大學(xué)》2007年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 翻譯增強(qiáng)子 omega序列 乙肝病毒表面抗原 人參愈傷組織

【摘要】： 我們?cè)谇捌诠ぷ髦薪⒘巳藚⒂鷤M織細(xì)胞表達(dá)乙肝病毒表面抗原(HBsAg)的植物表達(dá)系統(tǒng),并就提高表達(dá)進(jìn)行了培養(yǎng)條件的摸索。為了進(jìn)一步提高HBsAg的表達(dá),我們從表達(dá)載體的改造上做了一些探索,對(duì)調(diào)控HBsAg基因表達(dá)的啟動(dòng)子以及翻譯增強(qiáng)子進(jìn)行了改造,構(gòu)建了十二種不同的植物細(xì)胞表達(dá)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(zhuǎn)化農(nóng)桿菌LBA4404,再與人參愈傷組織細(xì)胞共培養(yǎng),經(jīng)G418篩選獲得新生抗性細(xì)胞,從而將HBsAg基因以及調(diào)控基因整合到人參細(xì)胞基因組中,經(jīng)過連續(xù)繼代培養(yǎng)選育出一株高產(chǎn)細(xì)胞系PBIo。 PBIo是由質(zhì)粒pBI/35Somega轉(zhuǎn)化的人參細(xì)胞選育而來,攜帶花椰菜花葉病毒CaMV 35S啟動(dòng)子以及與之串聯(lián)的煙草花葉病毒TMV(U1株)翻譯增強(qiáng)子omega序列。通過對(duì)表達(dá)載體的改造使HBsAg基因表達(dá)有顯著提高。在實(shí)驗(yàn)中還發(fā)現(xiàn),omega反義RNA序列也具有較強(qiáng)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(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2007
【分類號(hào)】：R392;Q943.2

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

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本文編號(hào)：1434884