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An alternative approach to the synthesis of a recombinant galanin-like peptide (GALP)

机译:合成重组甘丙肽样肽(GALP)的另一种方法

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摘要

GALP is a novel galanin-like peptide that has been isolated from porcine hypothalamus in our Research Division.nIn the present study, a large quantity of porcine GALP (pGALP) was synthesized with a high degree of purity usingnrecombinant DNA technology. A basic fibroblast growth factor (bFGF) mutein, CS23, was used as a fusion partnernand pGALP was released by chemical cleavage with cyanogen bromide (BrCN). Using various criteria such asnHPLC, SDS–PAGE, amino acid analysis, amino acid sequence, relative-molecular-mass measurement by liquidnsecondary-ion mass spectrometry, as well as biological activity, the purified molecule was shown to be pGALP andnwas found to have the same biological activity as a chemically synthesized standard. The pGALP obtained here willnbe useful for elucidation of its physiological role in vivo. Furthermore, CS23ML76, a newly prepared bFGF mutein,nwas found to be more appropriate than CS23 as a fusion partner in this procedure.
机译:GALP是一种新的甘丙肽样肽,已在我们的研究部门从猪下丘脑中分离出来。n在本研究中,使用重组DNA技术合成了大量具有高纯度的猪GALP(pGALP)。将碱性成纤维细胞生长因子(bFGF)突变蛋白CS23用作融合伴侣,并通过用溴化氰(BrCN)进行化学裂解来释放pGALP。使用nHPLC,SDS-PAGE,氨基酸分析,氨基酸序列,通过液相二次离子质谱法进行的相对分子质量测量以及生物学活性等各种标准,纯化的分子显示为pGALP,并且发现具有具有与化学合成标准品相同的生物活性。本文获得的pGALP将无法用于阐明其在体内的生理作用。此外,发现新制备的bFGF突变蛋白CS23ML76比CS23更适合作为该过程中的融合伴侣。

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