CONJUGATED PLASMID AND ITS PREPARATION

• Main Authors: NOJIRI CHIYUUHEI, YAMADA YUUJIROU, KATSUMATA KAZUKO
• Format: Patent
• Language: English
• Published: 25.02.1983
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; DERIVATIVES THEREOF; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; NUCLEIC ACIDS; NUCLEOSIDES; NUCLEOTIDES; ORGANIC CHEMISTRY; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; SUGARS; VINEGAR; WINE

PURPOSE:A stabilized, conjugated plasmid that is made by excising the base sequence concerning with the dissociation of a chain composed of plural multiplicative plasmids into each plasmid unit in the cells of a gram-positive bacterium, thus permitting stable cloning between different kinds of DNA. CONSTITUTION:A plurality of plamids, which are multiplicative in a gram-positive bacterium, especially in Bacillus subtilis, such as pUB110 and pC194 are introduced into cells of Bacillus subtilis, then, these cells are cultured to form a chain connecting these plasmid units (pCK1). This plasmid-connected chain is collected, treated with a restriction enzyme to excise the base sequence concerning with the dissociation of the chain into unit plasmids (assumed as the point near unit plasmids connections) and thet are connected again to prepare the objective conjugated plasmid. For example, EcoRI and Hind III are made to act on the above pCK1, then they are joined with T4 ligase to prepare a couple of conjugated plasmids of pCK2 of 3.7Md molecular weight and pCK3, 24Md molecular weight with 2 kinds of selective markers (Km and Cm).