Teng Chao-Yi, Wu Tzong-Yuan
Department of Bioscience Technology and Center for Nanotechnology, Chung Yuan Christian University, Chung-Li, Taiwan.
Biotechnol Lett. 2007 Jul;29(7):1019-24. doi: 10.1007/s10529-007-9349-y. Epub 2007 Mar 31.
The advantages of using traceable fluorescent protein (enhanced green fluorescent protein; EGFP) and a secretory alkaline phosphatase (SEAP) have been used to generate a reporter gene: the secretory fluorescent protein (SEFP). Sf21 cells, infected with the recombinant baculovirus containing the SEFP gene, revealed both traceable fluorescence and easily detectable alkaline phosphatase activity in the culture medium. The distribution of SEFP within the cells revealed that it was excluded from the nucleus, implying that the accumulation of SEFP in a secretory pathway, similar to that of the secretion signal-tagged FPs. Furthermore, the time- and dose-dependent release from the blockage of brefeldin A (BFA) confirmed that the secretion of SEFP was mediated by the secretion pathway and excluded leakage from viral infection. This SEFP reporter gene with traceable fluorescence and alkaline phosphatase activity may become a useful tool for studies on secretory protein production.
利用可追踪荧光蛋白（增强型绿色荧光蛋白；EGFP）和分泌性碱性磷酸酶（SEAP）的优势，已构建出一种报告基因：分泌性荧光蛋白（SEFP）。感染含SEFP基因重组杆状病毒的Sf21细胞，在培养基中呈现出可追踪的荧光以及易于检测的碱性磷酸酶活性。细胞内SEFP的分布显示其被排除在细胞核外，这意味着SEFP在分泌途径中的积累，类似于分泌信号标记的荧光蛋白。此外，布雷菲德菌素A（BFA）阻断后的时间和剂量依赖性释放证实，SEFP的分泌是由分泌途径介导的，排除了病毒感染导致的渗漏。这种具有可追踪荧光和碱性磷酸酶活性的SEFP报告基因可能成为研究分泌蛋白产生的有用工具。
