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Expression and Purification of Recombinant Human Apolipoprotein A-II in Pichia pastoris

机译:重组人载脂蛋白A-II在毕赤酵母中的表达和纯化

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摘要

Apolipoprotein A-II (ApoA-II) is the second most abundant protein constituent of high-density lipoprotein (HDL). The physiologic role of ApoA-II is poorly defined. ApoA-II may inhibit lecithin:cholesterol acyltransferase and cholesteryl-ester-transfer protein activities, but may increase the hepatic lipase activity. ApoA-II may also inhibit the hepatic cholesteryl uptake from HDL probably through the scavenger receptor class B type I depending pathway. Interpretation of data from transgenic and knockout mice of genes involved in lipoprotein metabolism has been often complicated as clinical implications because of species difference. So it is important to obtain human ApoA-II for further studies about its functions. In our studies, Pichia pastoris expression system was first used to express a high-level secreted recombinant human ApoA-II (rhApoA-II). We have cloned the cDNA encoding human ApoA-II and achieved its high-level secreting expression with a yield of 65 mg/L of yeast culture and the purification process was effective and easy to handle. The purified rhApoA-II can be used to further study its biological activities.
机译:载脂蛋白A-II(ApoA-II)是高密度脂蛋白(HDL)的第二大最丰富的蛋白质成分。 ApoA-II的生理作用定义不清。 ApoA-II可能抑制卵磷脂:胆固醇酰基转移酶和胆固醇酯转移蛋白的活性,但可能会增加肝脂肪酶的活性。 ApoA-II也可能通过I型清道夫受体依赖途径抑制HDL对肝胆固醇的吸收。来自脂蛋白代谢的基因的转基因和基因敲除小鼠的数据解释由于种属差异通常具有复杂的临床意义。因此,获得人类ApoA-II对其功能进行进一步研究很重要。在我们的研究中,巴斯德毕赤酵母表达系统首先用于表达高水平分泌的重组人ApoA-II(rhApoA-II)。我们已经克隆了编码人ApoA-II的cDNA,并以65μmg/ L的酵母培养物产量实现了其高水平的分泌表达,并且纯化过程有效且易于操作。纯化的rhApoA-II可用于进一步研究其生物学活性。

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