SECRETION OF RECOMBINANT RIBONUCLEASE-T1 INTO THE PERIPLASMIC SPACE OF ESCHERICHIA-COLI WITH THE AID OF THE SIGNAL PEPTIDE OF ALKALINE-PHOSPHATASE

被引：6

作者：

FUJIMURA, T ^{[1
]}

TANAKA, T ^{[1
]}

OHARA, K ^{[1
]}

MORIOKA, H ^{[1
]}

UESUGI, S ^{[1
]}

IKEHARA, M ^{[1
]}

NISHIKAWA, S ^{[1
]}

机构：

[1] OSAKA UNIV,FAC PHARMACEUT SCI,1-6 YAMADAOKA,SUITA,OSAKA 565,JAPAN

来源：

FEBS LETTERS | 1990年 / 265卷 / 1-2期

关键词：

Alkaline phosphatase; Recombinant protein; Ribonuclease T[!sub]1[!/sub; Secretion; Signal peptide;

D O I：

10.1016/0014-5793(90)80886-N

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The ribonuclease T1 (RNase T1) gene was ligated to a synthetic gene for the signal peptide of Escherichia coli alkaline phosphatase. When this fusion gene was expressed in E.Coli under the control of the trp promoter, active RNase T1 having the correct N-terminal sequence was secreted into the periplasmic space, indicating that the heterologous signal peptide had been cleaved off correctly. The enzyme could be readily purified from the periplasmic fraction with a yield of 1.8 mg from 1 liter culture. Adopting the same strategy, it was possible to produce a labile mutant of RNase T1 (Glu-58 → Ala mutant) in E. coli, the yield of the purified mutant enzyme being 2.0 mg from 1 liter culture. © 1990.

引用

页码：71 / 74

页数：4

共 16 条

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