本文選題：乙型肝炎病毒 + RNA剪接�。� 參考：《福建醫(yī)科大學(xué)》2011年碩士論文

【摘要】：HBV基因組剪接變異體(spliced variants of hepatitis B virus genomes)是一大類(lèi)由HBV前基因組RNA (pregenomic RNA, pgRNA)經(jīng)剪接并逆轉(zhuǎn)錄產(chǎn)生的亞基因組DNA。長(zhǎng)度為2.2 kb的HBV剪接變異體占80%以上,其中2447nt-489nt剪接變異體可編碼剪接特異蛋白HBSP (Hepatitis B spliced protein),并與病毒的持續(xù)性感染及致病性相關(guān),但其機(jī)制尚未完全闡明。 CytoTrap細(xì)胞質(zhì)酵母雙雜交系統(tǒng)將相互作用的蛋白錨定在細(xì)胞膜上、激活Ras信號(hào)通路,從而使溫度敏感突變株酵母細(xì)胞cdc25H能夠在37°C生長(zhǎng)。與經(jīng)典的酵母細(xì)胞核雙雜交相比,該系統(tǒng)更有助于篩查僅發(fā)生于細(xì)胞質(zhì)的蛋白相互作用;且更適用于誘餌蛋白定位于細(xì)胞質(zhì)并需要轉(zhuǎn)錄后翻譯修飾的情況。 本研究利用CytoTrap細(xì)胞質(zhì)酵母雙雜交系統(tǒng)篩選,獲得與HBSP相互作用的肝細(xì)胞蛋白—轉(zhuǎn)化生長(zhǎng)因子β1誘導(dǎo)蛋白(transforming growth factor beta 1 induced transcript 1 ,TGFβ1I1),并通過(guò)GST-pull down和內(nèi)源性免疫共沉淀在體內(nèi)外初步證實(shí),該蛋白能夠與HBSP相互作用。提示HBSP可能通過(guò)與TGFβ1I1相互作用,影響肝癌細(xì)胞侵襲能力、凋亡以及雄激素誘導(dǎo)的腫瘤發(fā)生,為深入闡明該剪接變異體致病性奠定基礎(chǔ)。
[Abstract]:The HBV genomic splicing variant (spliced variants of hepatitis B virus genomes) is a large class of sub genomic RNA (pregenomic RNA, pregenomic RNA), which is produced by splicing and reverse transcriptase, and is produced by the subgenome of 2.2. Is B spliced protein) is associated with persistent infection and pathogenicity of virus, but its mechanism has not yet been fully elucidated.
The CytoTrap cytoplasmic yeast two hybrid system anchors the interacting proteins on the cell membrane and activates the Ras signaling pathway so that the temperature sensitive mutant yeast cell cdc25H can grow at 37 degree C. Compared with the classical yeast cell nuclear double hybrid, the system is more helpful to screen the protein interaction only in the cytoplasm; and it is more suitable. The bait protein is located in the cytoplasm and needs post transcriptional modification.
In this study, CytoTrap cytoplasmic yeast two hybrid system was used to obtain the hepatocyte protein transforming growth factor beta 1 induced protein (transforming growth factor beta 1 induced transcript 1, TGF beta 1I1) interacting with HBSP, and it was preliminarily confirmed by GST-pull down and endogenous pestilence co precipitation in vivo and in vitro. BSP interaction suggests that HBSP may affect the invasiveness, apoptosis and androgen induced tumorigenesis of hepatoma cells through interaction with TGF beta 1I1, which lays the foundation for further elucidating the pathogenicity of the splicing variant.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R373

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 王海波;安學(xué)麗;張艷貞;王愛(ài)麗;李巧云;晏月明;;蛋白質(zhì)相互作用研究方法及其應(yīng)用[J];生物技術(shù)通報(bào);2006年S1期

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