| 摘要 |
NEW MATERIAL:Composite plasmid pHY310 having 3.1 megadalton molecular weight, and arrangement of recognition scission site of restricted enzyme shown by the figure. USE:Useful as a cloning vector of Escherichia coli and Bacillus subtilis synthesized from a plasmid of Streptococcus faecalis of Gram-positive bacteria and Escherichia coli of Gram-negative bacteria. PREPARATION:Composite plasmid pHY360 obtained by starting from plasmid pAMalpha1 containing a tetracycline-resistant gene range (Tc) of Streptococcus faecalis is scissored with restricted enzymes AccI and BamHI. Single stranded DNA parts existing both the ends of the scission part are treated with T4DNA polymerase and four kinds of deoxyribonucleotide-triphosphates, both the ends are processed into duplex flush ends, and both the ends of the scissored parts are connected through linker Hind III with ligase T4. |
