Manufactured by Yakult Co., Ltd.
>70% double-stranded covalently closed circular form I (RF I) DNA.
–20°C. Supplied with the host strain Bacillus subtilis ISW 1214 (Glycerol stock, Storage : –80°C)
10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA
Anagnostopoulos, C. & Spizizen, J. Requirements for Transformation in Bacillus Subtilus. J. Bacteriol. 81, 741–6 (1961).
Ikawa, S. et al. Chromosomal loci of genes controlling site-specific restriction endonucleases of Bacillus subtilis. Mol. Gen. Genet. 183, 1–6 (1981).
Ishiwa, H. & Shibahara, H. New shuttle vectors for Escherichia coli and Bacillus subtilis. II. Plasmid pHY300PLK, a multipurpose cloning vector with a polylinker, derived from pHY460. Japanese J. Genet. 60, 235–243 (1985).
Ishiwa, H. & Tsuchida, N. New shuttle vectors for Escherichia coli and Bacillus subtilis. I. Construction and characterization of plasmid pHY460 with twelve unique cloning sites. Gene 32, 129–34 (1984).
Sadaie, Y. & Kada, T. Formation of competent Bacillus subtilis cells. J. Bacteriol. 153, 813–21 (1983).
- E. coli: >2 × 105/µg DNA
- B. subtillis: >2 × 10/µg DNA (by electroporation)
Transformation efficiency of competent B. subtillis largely depends on the percentage of dimerized pHY300PLK vector. To increase transformation efficiency, first amplify pHY300PLK DNA with an E. coli recA+ strain.
Prior to electroporation of B. subtillis, amplify vector with an E.coli recA- strain to obtain monomeric pHY300PLK.