PURPOSE: To obtain the subject antibiotic in high yield by screening a pNA gene cluster encoding for the biosynthetic pathway of tetracycline, chlorotetracycline or an analogue thereof produced by microorganisms, allowing the screened DNA gene cluster to express and controlling the biosynthetic pathway.
CONSTITUTION: The objective antibiotics or analogues thereof are obtained in high yield by culturing Streptomyces aureofaciens ATCC13899 in a medium, gathering the cultured microbes by centrifugation, carrying out protease treatment by dispersing the gathered microbes in a buffer, carrying out lysis of the treated microbes by adding a surfactant, forming a library by treating the microbe after the lysis with a restriction enzyme and combining the treated microbe with a cloning vector, screening a resistant transformant including a DNA gene cluster encoding for the biosynthetic pathway of tetracycline or chlorotetracycline, or a part thereof, extracting the gene cluster of the positive strain, combining the extracted gene cluster with an expression vector by treating with the restriction enzyme, transforming a host cell by using the combined expression vector, and culturing the transformed host cell.
MAIKERU JIEI RAIAN
NANSHII SUTORASHII
