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

[1] CLONING AND EXPRESSION OF A BACILLUS-COAGULANS AMYLASE GENE IN ESCHERICHIA-COLI
CORNELIS, P
DIGNEFFE, C
WILLEMOT, K
[J]. MOLECULAR AND GENERAL GENETICS, 1982, 186 (04): : 507 - 511
[2] CONFORMATIONAL PROPERTIES OF THE GUANINE-BINDING SITE OF RIBONUCLEASE-T1 INFERRED FROM THE X-RAY STRUCTURE AND PROTEIN ENGINEERING
HAKOSHIMA, T
TODA, S
SUGIO, S
TOMITA, K
NISHIKAWA, S
MORIOKA, H
FUCHIMURA, K
KIMURA, T
UESUGI, S
OHTSUKA, E
IKEHARA, M
[J]. PROTEIN ENGINEERING, 1988, 2 (01): : 55 - 61
[3] INQUIRIES INTO THE STRUCTURE-FUNCTION RELATIONSHIP OF RIBONUCLEASE-T1 USING CHEMICALLY SYNTHESIZED CODING SEQUENCES
IKEHARA, M
OHTSUKA, E
TOKUNAGA, T
NISHIKAWA, S
UESUGI, S
TANAKA, T
AOYAMA, Y
KIKYODANI, S
FUJIMOTO, K
YANASE, K
FUCHIMURA, K
MORIOKA, H
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (13) : 4695 - 4699
[4] SYNTHESIS OF A GENE FOR HUMAN GROWTH-HORMONE AND ITS EXPRESSION IN ESCHERICHIA-COLI
IKEHARA, M
OHTSUKA, E
TOKUNAGA, T
TANIYAMA, Y
IWAI, S
KITANO, K
MIYAMOTO, S
OHGI, T
SAKURAGAWA, Y
FUJIYAMA, K
IKARI, T
KOBAYASHI, M
MIYAKE, T
SHIBAHARA, S
ONO, A
UEDA, T
TANAKA, T
BABA, H
MIKI, T
SAKURAI, A
OISHI, T
CHISAKA, O
MATSUBARA, K
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1984, 81 (19): : 5956 - 5960
[5] CONSTRUCTION OF AN EXCRETION VECTOR AND EXTRACELLULAR PRODUCTION OF HUMAN GROWTH-HORMONE FROM ESCHERICHIA-COLI
KATO, C
KOBAYASHI, T
KUDO, T
FURUSATO, T
MURAKAMI, Y
TANAKA, T
BABA, H
OISHI, T
OHTSUKA, E
IKEHARA, M
YANAGIDA, T
KATO, H
MORIYAMA, S
HORIKOSHI, K
[J]. GENE, 1987, 54 (2-3) : 197 - 202
[6] THE NUCLEOTIDE-SEQUENCE OF THE PROMOTER AND THE AMINO-TERMINAL REGION OF ALKALINE-PHOSPHATASE STRUCTURAL GENE (PHOA) OF ESCHERICHIA-COLI
KIKUCHI, Y
YODA, K
YAMASAKI, M
TAMURA, G
[J]. NUCLEIC ACIDS RESEARCH, 1981, 9 (21) : 5671 - 5678
[7] CODON USAGE TABULATED FROM THE GENBANK GENETIC SEQUENCE DATA
MARUYAMA, T
GOJOBORI, T
AOTA, S
IKEMURA, T
[J]. NUCLEIC ACIDS RESEARCH, 1986, 14 : R151 - R197
[8] 2 HISTIDINE-RESIDUES ARE ESSENTIAL FOR RIBONUCLEASE-T1 ACTIVITY AS IS THE CASE FOR RIBONUCLEASE-A
NISHIKAWA, S
MORIOKA, H
KIM, HJ
FUCHIMURA, K
TANAKA, T
UESUGI, S
HAKOSHIMA, T
TOMITA, K
OHTSUKA, E
IKEHARA, M
[J]. BIOCHEMISTRY, 1987, 26 (26) : 8620 - 8624
[9] INCREASE IN NUCLEOLYTIC ACTIVITY OF RIBONUCLEASE-T1 BY SUBSTITUTION OF TRYPTOPHAN 45 FOR TYROSINE 45
NISHIKAWA, S
MORIOKA, H
KIMURA, T
UEDA, Y
TANAKA, T
UESUGI, S
HAKOSHIMA, T
TOMITA, K
OHTSUKA, E
IKEHARA, M
[J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1988, 173 (02): : 389 - 394
[10] GLU-46 OF RIBONUCLEASE-T1 IS AN ESSENTIAL RESIDUE FOR THE RECOGNITION OF GUANINE BASE
NISHIKAWA, S
KIMURA, T
MORIOKA, H
UESUGI, S
HAKOSHIMA, T
TOMITA, K
OHTSUKA, E
IKEHARA, M
[J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1988, 150 (01) : 68 - 74

← 1 2 →