Development of an efficient bioreactor system for delivering foreign proteins secreted from liver into eggs with a vitellogenin signal in medaka Oryzias latipes
Development of an efficient bioreactor system for delivering foreign proteins secreted from liver into eggs with a vitellogenin signal in medaka Oryzias latipes
复制标题
开发一种有效的生物反应器系统，用于将肝脏分泌的外源蛋白质传递到带有卵黄原蛋白信号的青鳉鱼卵中
DOI：
10.1007/s12562-019-01320-4
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发表时间：
2019
影响因子：
1.9
通讯作者：
Masato Kinoshita
中科院分区：
文献类型：
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作者：
Yu Murakami; Tomohisa Horibe;Masato Kinoshita
在这项研究中，我们开发了一种新的生物反应器系统，以青鳉鱼Oryzias latipes与援助的部分序列的卵黄蛋白原（Vtg）的鸡蛋中提供和积累外源蛋白。在硬骨鱼中，Vtg是肝脏产生的蛋黄蛋白的前体，它被分泌到血液中，然后被吸收到鸡蛋中。我们通过计算机模拟预测了Vtg的一个可能区域（Vtg信号），该区域介导蛋白质从肝脏运输到鸡蛋中。然后，我们建立了两个转基因株系，在肝脏中表达融合蛋白，包括Vtg信号和每个报告基因，增强型绿色荧光蛋白（EGFP）或荧光素酶（LUC）融合的EGFP，由肝脏特异性chgH（chgH）启动子驱动。从转基因雌性产卵的受精卵中检测到每个报告信号，表明蛋白质成功转运到具有Vtg信号的卵中。这是第一份证明Vtg信号具有将外源蛋白递送到鸡蛋中的能力的报告。由于Vtg是大多数卵生生物中高度保守的蛋白质，因此我们的研究结果有望建立在广泛生物中可行的生物反应器系统。
In this study, we developed a novel bioreactor system to deliver and accumulate foreign proteins in eggs using medaka fishOryzias latipeswith the aid of a partial sequence of vitellogenin (Vtg). In teleost fish, Vtg, the hepatically generated precursor of egg yolk proteins, is secreted into the bloodstream and then taken up into eggs. We predicted in silico a probable region (Vtg signal) of Vtg that mediates transportation of proteins from the liver into eggs. Then, we established two transgenic lines expressing the fused proteins including the Vtg signal and each reporter gene,enhanced green fluorescent protein(EGFP) or fireflyluciferase(LUC)-fusedEGFP, in the liver driven by a liver-specificchoriogeninH(chgH) promoter. Each reporter signal was detected from the fertilized eggs spawned by the transgenic females, showing successful transportation of the proteins into the eggs with the Vtg signal. This is the first report demonstrating that the Vtg signal has capability to deliver exogenous proteins into eggs. Because Vtg is a highly conserved protein among most of oviparous organisms, our findings hold promise for establishing bioreactor systems viable in a wide range of organisms.