Shotgun cloning of beta-amylase gene = 베타 아밀라제 유전자의 Shotgun cloning

Chromosomal DNA was isolated from Bacillus megaterium and plasmid pUB110 from Bacillus subtilis BD630. The chromosomal DNA was digested with xbaI and the 1.5 to 2.5 kilo-base fragments were isolated with DEAE paper and ligated to the plasmid pUB110 digested with xbaI and dephosphorylated with bacterial alkaline phosphatase. Competent Bacillus subtilis cells were prepared by treatment with 50 mM CaCl$_2$ solution and were transformed with the hybrid plasmids. And the kanamycin resistence transformants were screened for the production of beta-amylase. Among the 2,000 transformant colonies the 910th colony produced high amount of beta -amylase. The recombinant plasmid pBMA910 harbouring cell showed amylase activity about 250-fold higher than that of the transformation recipient cell Bacillus subtilis BD170. The fact that this enzyme is excreted so much shows me the possibility to use the plasmid pBMA910 as a new cloning vector for secretion of foreign gene products.
Advisors
Kim, Hyoung-Manresearcher김형만researcher
Publisher
한국과학기술원
Issue Date
1984
Identifier
64058/325007 / 000821223
Language
eng
Description

학위논문(석사) - 한국과학기술원 : 생물공학과, 1984.2, [ 52 p. ]

URI
http://hdl.handle.net/10203/28178
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=64058&flag=t
Appears in Collection
BS-Theses_Master(석사논문)
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