Krasevec N, van den Hondel C A, Komel R
National Institute of Chemistry, Ljubljana, Slovenia.
Pflugers Arch. 2000;439(3 Suppl):R84-6.
A gene-fusion expression strategy was applied for the heterologous expression of hTNF-alpha in A. niger AB1.13. The TNF-alpha gene was fused with the A. niger glucoamylase GII form as a carrier-gene, behind its transcription control and secretion signal. The protein was expressed in the cells in the form of a glucoamylase-fusion protein, but was not present in the culture medium. From the expression of two hTNF-alpha analogues, LK 811 (Cys95/148) and LK 802 (Cys95/148, His107/108) respectively, we concluded that oligomerisation was not the critical point for secretion of hTNF-alpha in A. niger, but more probably improper folding already at the stage of monomer formation, or even incorrect processing of the molecule during the secretion pathway.
一种基因融合表达策略被应用于在黑曲霉AB1.13中异源表达人肿瘤坏死因子-α（hTNF-α）。TNF-α基因与黑曲霉糖化酶GII形式作为载体基因融合，位于其转录控制和分泌信号之后。该蛋白以糖化酶融合蛋白的形式在细胞中表达，但不存在于培养基中。从分别表达的两种hTNF-α类似物LK 811（Cys95/148）和LK 802（Cys95/148，His107/108）中，我们得出结论，寡聚化不是黑曲霉中hTNF-α分泌的关键点，更可能是在单体形成阶段就已经存在折叠不当，甚至在分泌途径中分子的加工不正确。
