1986 Volume 50 Issue 11 Pages 2845-2852
The Rsal fragment (750 base pairs) containing the entire early sporulation gene spo0F of Bacillus subtilis was inserted in the downstream region of the PR promoter of the expression vector, pEBR-151. The recombinant plasmid thus obtained was introduced into Escherichia coli HB101 and the synthesis of the spo0F gene product was induced by a temperature shift up. After induction for 7 hr, a protein of molecular weight 14, 000 (14 K protein) was overproduced to about 9% of the total cellular protein. The 14K protein was purified to 94% purity by four steps of column chromatography. Deletion analysis and the sequence determination of the NH2-terminal amino acid residues of the purified 14 K protein confirmed that the 14 K protein is the spo0F gene product.
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