人二氢蝶啶还原酶（EC 1.6.99.7）在大肠杆菌中的高水平表达，无N端氨基酸保护

Armarego W L, Cotton R G, Dahl H H, Dixon N E

Division of Biochemistry and Molecular Biology, John Curtin School of Medical Research, Australian National University, Canberra.

Biochem J. 1989 Jul 1;261(1):265-8. doi: 10.1042/bj2610265.

The cDNA coding for human dihydropteridine reductase [Dahl, Hutchinson, McAdam, Wake, Morgan & Cotton (1987) Nucleic Acids Res. 15, 1921-1936] was inserted downstream of tandem bacteriophage lambda PR and PL promoters in Escherichia coli vector pCE30. Since pCE30 also expresses the lambda c1857ts gene, transcription may be controlled by variation of temperature. The recombinant plasmid in an E. coli K12 strain grown at 30 degrees C, then at 45 degrees C, directed the synthesis of dihydropteridine reductase to very high levels. The protein was soluble, at least as active as the authentic human enzyme, and lacked the N-terminal amino acid protection.

编码人二氢蝶啶还原酶的cDNA[达尔、哈钦森、麦克亚当、韦克、摩根和科顿（1987年）《核酸研究》15卷，1921 - 1936页]被插入大肠杆菌载体pCE30中串联的噬菌体λ PR和PL启动子的下游。由于pCE30也表达λ c1857ts基因，转录可通过温度变化来控制。在30℃培养然后在45℃培养的大肠杆菌K12菌株中的重组质粒，指导合成了非常高水平的二氢蝶啶还原酶。该蛋白质是可溶的，至少与天然人酶一样有活性，并且缺乏N端氨基酸保护。