双启动子表达载体转化CHO、HEK-293和vero细胞株中丁基胆碱酯酶的表达和活性评价

Q3 Biochemistry, Genetics and Molecular Biology Journal of Cellular Biotechnology Pub Date : 2022-02-25 DOI:10.3233/jcb-210042
Vida Mirzaie, Touba Eslaminejad, H. Babaei, S. Nematollahi-Mahani
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引用次数: 0

摘要

背景:丁酰胆碱酯酶(BChE)是一种治疗药物,其作为重组蛋白的生产是生物技术中的一个重要问题。这方面的一个亮点是选择最好的宿主细胞和质粒。目的:本研究的目的是使用双启动子载体评估Vero、HEK-293和CHO细胞系中丁酰胆碱酯酶的产生。材料和方法:将双启动子构建体(pBudCE双BChE)转染到分为三个实验组的细胞系中(pBudCE双BChE、pCMV和阴性对照)。在不同时间评估BChE基因表达和酶活性。结果:pBudCE双BChE组三种细胞系均表现出较高的基因表达水平。CHO细胞BChE酶活性第6天下降,第9天上升。在HEK-293细胞中,从第六天到第九天它有下降的趋势,而在Vero细胞中,第九天的水平最高。结论:pBudCE双BChE和pCMV组在HEK-293细胞中的差异更为明显,该细胞的BChE基因表达水平高于其他细胞,而CHO细胞的BCh E酶活性水平更高。
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Evaluation of the butyrylcholinesterase expression and activity in CHO, HEK-293 and vero cell lines transformed by dual promoter expression vector
BACKGROUND: Butyrylcholineesterase (BChE) is a therapeutic drug and its producing as a recombinant protein is an essential issue in biotechnology. One of the highlights in this regard is choosing the best host cells and plasmids. OBJECTIVES: The aim of this study is to evaluate the production of butyrylcholinesterase in Vero, HEK-293, and CHO cell lines using a dual promoter vector. MATERIAL AND METHODS: The dual-promoter construction (pBudCE dual BChE) was transfected into cell lines categorized in three experimental groups (pBudCE dual BChE, pCMV and negative control). BChE gene expression and enzyme activity was evaluated at different times. RESULTS: All three cell lines showed higher gene expression level in pBudCE dual BChE group. BChE enzyme activity level of this group in CHO cells decreased in sixth day and increased in ninth day. In HEK-293 cells it has a downward trend from sixth to ninth day and in Vero cells its level in the ninth day was the highest. CONCLUSION: The difference of pBudCE dual BChE and pCMV groups was more pronounced in the HEK-293 cell and the BChE gene expression level of this cells was higher than the others while, CHO cells showed higher level of BChE enzyme activity.
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来源期刊
Journal of Cellular Biotechnology
Journal of Cellular Biotechnology Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
0.70
自引率
0.00%
发文量
13
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