Overproduction and purification of the CepA protein from Lactococcus lactis

• Autorzy: Kowalczyk M. , Bardowski J.
• Języki publikacji: EN

Abstrakty

EN

In this work we present cloning and overexpression of lactococcal CcpA protein in Escherichia coli Xllblue strain as a fusion with 6xHis tag. A high yield of the CcpA protein was obtained when the cells were cultured in liquid medium LB with 100 ug/ml ampicillin at 37°C and subsequently for 4 h after induction by IPTG. The proce­dure let us obtain 5 mg of homogenous CcpA protein. Glutaraldehyde crosslinking analysis indicated the formation of dimer or tetramer forms of the CcpA protein.

Słowa kluczowe

EN

CepA protein; overproduction; cloning; Lactococcus lactis; protein; purification; Escherichia coli; overexpression; dimer

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2003
• Tom: 50
• Numer: 2
• Opis fizyczny: p.455-459,fig.

Twórcy

autor

Kowalczyk M.

• Polish Academy of Sciences, A.Pawinskiego 5a, 02-106 Warsaw, Poland

autor

Bardowski J.

Bibliografia

• Aleksandrzak T, Kowalczyk M, Kok J, Bardowski J. (2000) Regulation of carbon catabolism in Lactococcus lactis. In Food Biotechnology, Bielecki S, Tramper J, Polak J. eds, 17: 61-6. Elsevier Science B.V.
• Bradford MM. (1976) AnalBiochem.; 72: 248-54.
• Deutscher J, Reizer J, Fischer C, Galinier A, Saier MH, Jr, Steinmetz M. (1994) J Bacteriol.; 176: 3336-44.
• Deutscher J, Kuster E, Bergstedt U, Charrier V, Hillen W. (1995) Mol Microbiol.; 15:1049-53.
• Gibson TJ. (1984) Studies on the Epstein-Barr virus genome. Ph.D. Thesis.; Cambridge University Press.
• Gosseringer R, Kuster E, Galinier A, Deutscher J, Hillen W. (1997) J Mol Biol.; 266: 665-76.
• Guedon E, Jamet E, Renault P. (2002) Antonie Van Leeuwenhoek.; 82: 93-112.
• Mahr K, Esteban CD, Hillen W, Titgemeyer F, Perez-Martinez G. (2002) J Mol Microbiol Biotechnol., 4: 489-94.
• Miwa Y, Saikawa M, Fujita Y. (1994) Microbiology.; 140: 2567-75.
• Sambrook J, Fritsch EF, Maniatis T. (1989) Molecular cloning a laboratory manual. Cold Spring Harbor Laboratory Press.
• Sanger F, Nicklen S, Coulson AR. (1977) Proc Natl Acad Sci U S A.; 74: 5463-7. Schick J, Weber B, Klein JR, Henrich B. (1999) Microbiology.; 145: 3147-54.
• Tebbe J, Orth P, Kuster-Schock EK, Hillen W, Saenger W, Hinrichs W. (2000) Acta Crystallogr D Biol Crystallogr.; 56: 67-9.
• Titgemeyer F, Hillen W. (2002) Antonie Van Leeuwenhoek.; 82: 59-71. Weickert MJ, Chambliss GJ. (1990) Proc Natl Acad Sci US A.; 87: 6238-42.