>70% double-stranded covalently closed circular form I (RF I) DNA.
- 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.
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-Bacillus Shuttle Vector pHY300PLK
–20°C. Supplied with the host strain Bacillus subtilis ISW 1214 (Glycerol stock, Storage : –80°C)
E. coli-Bacillus Shuttle Vector | pHY300PLK
10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA
The E.coli-Bacillus Shuttle Vector pHY300PLK is constructed from the E.coli-derived plasmid pACYC177 and the Streptococcus faecalis-derived plasmid pAMa1; as a result, the vector can be used to transform both E. coli and Bacillus subtillis. The E. coli-Bacillus Shuttle Vector DNA contains both the ampicillin resistance and tetracycline resistance genes as selective markers; while both of these resistance genes can be expressed in E. coli, only the tetracycline resistance gene can be expressed in B. subtillis. The pHY300PLK vector also contains a single cleavage site for restriction enzymes Bal I, BamH I, Ban I, Bgl I, Bgl II, Bst P I, EcoR I, EcoR V, Hind III, Hpa I, Sal I, Sma I, Pvu I and Xba I.
Manufactured by Yakult Co., Ltd.