Description

This DNA is a shuttle vector which is constructed with a plasmid pACYC177 of E. coli and DNA derived from a plasmid pAMα1 of Streptococcus faecalis. This DNA can transform both E. coli and Bacillus subtillis. Since this DNA has ampicillin resistant gene and tetracycline resistant gene as selective marker, both genes can be expressed in E. coli, but only tetracycline resistant gene can be expressed in B. subtillis. The vector contains a single cleavage site for 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.

Storage

-20°C
Supplied with Host strain: Bacillus subtilis ISW 1214 (Glycerol stock, Storage : -80°C)

Concentration

300-800 µg/ml

Chain length

4,870 bp

DNA Sequences of PHY300PLK DNA

pHY300PLK DNA

Purity

Contains over 70% double-strand covalently closed circular form I (RF I).

Transformation efficiency

E. coli > 2 × 10⁵/µg DNA
B. subtillis > 1 × 10²/µg DNA (electroporation method)

Note

Transformation efficiency using competent cells by plasmid of B. subtillis largely depends on the contents of dimer. Therefore, to increase frequency of transformation, this DNA must be amplified in E. coli recA⁺ strain prior to transformation. In case of electroporation, use of the monomer plasmid is recommended.

Form

10 mM Tris-HCl (pH7.4), 0.1 mM EDTA

Restriction map of pHY300PLK DNA