Abstract:

Based on the flanking sequence of the agarase gene of FG4 strain, the recombinant plasmid and vector were constructed to obtain the engineering strain which could express the agarase efficiently. In this study, the flanking sequence of FG4 agarase gene is obtained by chromosome walking. The cloned gene is recombined with the expression vector pET-22b （+）and transferred into the recipient BL21. After the successful construction of the expressed strain, the expression of agarase is induced by IPTG and the optimal fermentation conditions of the expressed strain are explored. This strain ensures the catalytic activity of agarase and lays the foundation for the agarase-based pharmaceutical industry, daily chemical industry and food industry. The cloning and expression of agarase gene from Microbulbifers sp. FG4 strain of Microbulbifers provides theoretical support for agarase application in different fields.

Key words: Micrcbulbifer sp., agarase, chromosome walking, cloning and expression