活性牛末端脱氧核苷酸转移酶(TdT)在大肠杆菌中的产生

Wee Liang Kuan, J. Joy, Ng Fui Mee, Kwek Zekui Perlyn, Then Siew Wen, T. Nguen, Joanne James, E. Chai, H. Flotow, Sharon C. Crasta, Kelvin Chua, Ng Sok Peng, Jeffrey Hill
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引用次数: 0

摘要

合成了牛末端脱氧核苷酸转移酶(TdT)基因,并将其克隆到表达载体中,在大肠杆菌中表达。研究了不同培养和诱导条件对重组蛋白性质的影响。这导致了重组牛TdT在大肠杆菌中的表达。纯化和鉴定后,酶的活性在细胞凋亡的生物测定中被评估。本报告中描述的过程使高通量应用的TdT经济生产成为可能。
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Generation of Active Bovine Terminal Deoxynucleotidyl Transferase (TdT) in E.coli
A synthetic gene encoding bovine terminal deoxynucleotidyl transferase (TdT) was generated, cloned into an expression vector and expressed in E.coli. The effects of altering culture and induction conditions on the nature of recombinant protein production were investigated. This led to the expression of active recombinant bovine TdT in E.coli. After purification and characterisation, the activity of the enzyme was assessed in a biological assay for apoptosis. The process described in this report enables the economical production of TdT for high throughput applications.
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Biochemistry Insights
Biochemistry Insights BIOCHEMISTRY & MOLECULAR BIOLOGY-
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