Amplification and purification of plasmid-encoded thioredoxin from Escherichia coli K12.
作者： C A Lunn , S Kathju , B J Wallace , S R Kushner , V Pigiet
DOI: 10.1016/S0021-9258(18)90987-7
关键词:
摘要: The thioredoxin gene (trxA) from Escherichia coli K12 has been cloned on a 3-kilobase pair PvuII fragment in derivative of pBR325 (pBHK8). Thioredoxin protein production was amplified 150-200-fold strain containing pBHK8 (SK3981), with the greatest increase/cell observed after cultures reached stationary phase. A simple purification procedure, involving DEAE and AcA-54 column chromatography, yielded homogeneous approximately 70% yield. high amplification these cells (i.e. 10(6) copies/cell representing 40% total cell protein) approaches maximum yields seen genetically constructed cloning vehicles (Bernard, H.U., Helinski, D.R. (1980) Genetic Engineering (Setlow, J. K., Hollaender, A., eds) Vol. 2, pp. 133-167, Plenum Press, New York). This tremendous overproduction is attributed to plasmid copy number SK3981 (1700/cell). These results suggest role for DNA replication.
sci-hub.st 参考文章(26)
V. Pigiet, R.R. Conley, Isolation and characterization of phosphothioredoxin from Excherichia coli. Journal of Biological Chemistry. ,vol. 253, pp. 1910- 1920 ,(1978) , 10.1016/S0021-9258(19)62335-5
Tadashi Asahi, Robert S. Bandurski, Lloyd G. Wilson, Yeast sulfate-reducing system. II. Enzymatic reduction of protein disulfide. Journal of Biological Chemistry. ,vol. 236, pp. 1830- 1835 ,(1961) , 10.1016/S0021-9258(19)63311-9
Simon Black, Eileen M. Harte, Blondel Hudson, Leonard Wartofsky, A Specific Enzymatic Reduction of l(-) Methionine Sulfoxide and a Related Nonspecific Reduction of Disulfides Journal of Biological Chemistry. ,vol. 235, pp. 2910- 2916 ,(1960) , 10.1016/S0021-9258(18)64561-2
A Holmgren, Glutathione-dependent synthesis of deoxyribonucleotides. Purification and characterization of glutaredoxin from Escherichia coli. Journal of Biological Chemistry. ,vol. 254, pp. 3664- 3671 ,(1979) , 10.1016/S0021-9258(18)50813-9
C A Lunn, V P Pigiet, Localization of thioredoxin from Escherichia coli in an osmotically sensitive compartment. Journal of Biological Chemistry. ,vol. 257, pp. 11424- 11430 ,(1982) , 10.1016/S0021-9258(18)33776-1
A. Holmgren, I. Ohlsson, M.L. Grankvist, Thiroedoxin from Escherichia coli. Radioimmunological and enzymatic determinations in wild type cells and mutants defective in phage T7 DNA replication. Journal of Biological Chemistry. ,vol. 253, pp. 430- 436 ,(1978) , 10.1016/S0021-9258(17)38227-3
C.H.W. Hirs, The oxidation of ribonuclease with performic acid. Journal of Biological Chemistry. ,vol. 219, pp. 611- 621 ,(1956) , 10.1016/S0021-9258(18)65721-7
M R Krauss, R M Benbow, Polyoma virus minichromosomes: associated DNA molecules. Journal of Virology. ,vol. 38, pp. 815- 825 ,(1981) , 10.1128/JVI.38.3.815-825.1981
K. Ito, T. Date, W. Wickner, Synthesis, assembly into the cytoplasmic membrane, and proteolytic processing of the precursor of coliphage M13 coat protein. Journal of Biological Chemistry. ,vol. 255, pp. 2123- 2130 ,(1980) , 10.1016/S0021-9258(19)86002-7
G.B. Kallis, A. Holmgren, Differential reactivity of the functional sulfhydryl groups of cysteine-32 and cysteine-35 present in the reduced form of thioredoxin from Escherichia coli. Journal of Biological Chemistry. ,vol. 255, pp. 10261- 10265 ,(1980) , 10.1016/S0021-9258(19)70458-X