1994
DOI: 10.1006/prep.1994.1018
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Purification Procedure for Bacterial Translational Initiation Factors IF2 and IF3

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Cited by 35 publications
(23 citation statements)
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“…IF2 was isolated by using the method described in ref. 24. A mRNA transcript with the sequence GGGAAUUCGGGCCCUU-GUUAACAAUUAAGGAGGUAUACUAUGAAUGCAAUA-AAUAACUGCAG(A) 21 (25) was prepared by in vitro transcription and purified according to ref.…”
Section: Methodsmentioning
confidence: 99%
“…IF2 was isolated by using the method described in ref. 24. A mRNA transcript with the sequence GGGAAUUCGGGCCCUU-GUUAACAAUUAAGGAGGUAUACUAUGAAUGCAAUA-AAUAACUGCAG(A) 21 (25) was prepared by in vitro transcription and purified according to ref.…”
Section: Methodsmentioning
confidence: 99%
“…Cells were opened by addition of lysozyme (5 mg͞g wet cells), sodium deoxycholate (12.5 mg͞g), and DNase I in buffer C (20 mM Tris⅐HCl, pH 7.7͞60 mM NH 4 Cl͞10 mM MgCl 2 ͞5 mM 2-mercaptoethanol͞0.1 mM PMSF). To wash initiation factors off the ribosomes, 1 M KCl was added to the S30 fraction and incubated for 10 min, and the ribosomes were removed by centrifugation at 100,000 ϫ g. From the S100 extract, initiation factors were purified to homogeneity by Fast-Flow LC on S-Sepharose by using a gradient of 0.2-0.8 M KCl in 50 mM Mops, pH 6.4͞0.25 mM MgCl 2 ͞0.05 mM EDTA͞5 mM 2-mercaptoethanol͞0.1 mM PMSF (14).…”
Section: Methodsmentioning
confidence: 99%
“…KsgAE66A was eluted with the same buffer containing 250 mM imidazole. IF-3 was purified as previously described (Soffientini et al 1994). Cells were lysed in 20 mM Tris-HCl, pH 7.7, 60 mM NH 4 Cl, 10 mM MgCl 2 , 10% glycerol, 5 mM BME, 0.1 mM PMSF, and 0.1 mM benzamidine and the cleared lysate loaded onto a HiTrap Ni +2 column.…”
Section: Expression and Purification Of Mutant E Coli Ksga And If3mentioning
confidence: 99%