α-1,3 Glucanase 유전자의 Escherichia coli에서의 cloning 및 발현
Cloning and Expression of a -1,3 glucanase from Streptomyces Y9343 into Escherichia coli
윤정원, 홍성열,
소속 상세정보
윤정원 ( ) - 수원대학교 유전공학과
홍성열 ( ) - 수원대학교 유전공학과
KMID : 0355419960200040531
Abstract
Insoluble glucan produced by Streptococcus mutans and Streptococcus sanguis has been implicated as causative ag6nts of dental caries, which is very hardly degradable by common dextranasee because which contained insoluble a -1,3
´
glucosidic bond. a -1,3 glucanase produced by Stretomyces Y9343 was able to degrade insoluble glucan but it´s productivity was low. The a -1,3 glucanase gene from Streptomyces Y9343 was cloned into Escherichia coli JM109 with pUC19. Genomic DNA from Streptomyces Y9343 was partially digested with the restriction enzyme Hind III and elude fragment of 2.4 Kb size from genomic DNA fragments. This 2.4 Kb DNA ligated into Hind III digested pUC19 for transformation of E. coil M109. The primary selection of transformants was made by observation of white colonies of ampicillin-resistant clone by pUC19 containing ampicillin-resistance gene and lacZ gene. From the selected transformant, poistive clones of a -1,3 glucanase gene were detected as the clear zones on a modified Czapeck-Dox agar medium containing the insoluble glucan as a sole carbon source. This transformant possessed a plasmid, deginated pYH2, which contained the vector DNA and a 2.4 Kb Hind III insertion fragment, which was encoded a -1,3 glucanase and was named Escherichia coli TH722.
The - a 1,3 glucanase produced by transformant E. coli TH722 are going on for optimum conditions of culture and a protocol of purification.
키워드
α-1;3 Glucanase 유전자;Escherichia coli;cloning 및 발현
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