Ketol-acid reductoisomerase-producing Escherichia coli bacterial culture

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Catalogue Number	160657
Antigen/Gene or Protein Targets	Candida albicans Ketol-acid reductoisomerase
Bacterial Resistance	Carbenicillin and chloramphenicol
Synonyms	Ilv5
In vitro applications	Recombinant protein expression
Notes	For expression of recombinant proteins: E. coli cells were cultured in 5 ml LB broth supplemented with 50 μg/ml carbenicillin and 34 μg/ml chloramphenicol at 37°C and 120 rpm overnight. The next day, 250 μl of the culture was added to a new flask containing 25 ml LB with antibiotics and incubated at 37°C and 120 rpm for 24 h. Then, the culture was transferred into 500 ml LB broth, without antibiotics, and was grown for 3h 30 min at 37°C at 120 rpm. To induce the transcription of the inserted target gene, IPTG to a final concentration of 1 mM was added and the cultures were further incubated for 4 h in the same conditions. Lastly, the cell pellets were collected by centrifugation and stored at -80°C until further use. Recombinant proteins were obtained disrupting the pellet by sonication with pulses of 2 seconds for 5 min and 40 of amplitude.
Research Area	Bacteriology

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Andoni Ramirez Garcia

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	University of the Basque Country, EHU
	13 reagents listed on Ximbio

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