Development of the fusion technique with protplast of lactobacillus casei = 유산균에 있어서 프로토플라스트를 이용한 세포융합기술 개발에 관한 연구

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Lactobacillus. casei ATCC 7469 was successfully converted to protoplast by treating with endo-N-acetyl muramidase(4ug/ml) in 0.9M sucrose phosphate buffer. For full hydrolysis of cell wall, not only treatment of this enzyme but high concentration of sucrose and cold shock were necessary. 5mM of $MgCl_2$ has enhanced the stability of protoplasting cell. Under these conditions, the obtained protoplasts were taken a photography. The cell wall regeneration of protoplast was more effective on gelatin induced regeneration media than soft overlay method. The concentration of gelatin was optimal at 2.5\%. The frequence of regeneration was 6\% with treating of the enzyme (4ug/ml) for 20min. To get genetic marker for detection of fusiont, the mutagenesis with UV irradiation and NTG treatment was carried out. And the streptomycin resistant(400ug/ml) and rifampicin resistant(10ug/ml) mutants were isolated and maintained the stability for several transfer. In use of these marker, protoplast fusion could be carried out. PEG(polyethyleneglycol) used as a fusogen. Molecular weight of PEG was optimal at 6,000 and it``s concentration was 50\%. As a result of this experiment, $10^{-5}$ of frequence of fusion could be obtained and it is higher than frequence of spontaneous mutaion.
Advisors
Kim, Jung-Hoeresearcher김정회researcher
Description
한국과학기술원 : 생물공학과,
Publisher
한국과학기술원
Issue Date
1984
Identifier
64041/325007 / 000821004
Language
eng
Description

학위논문(석사) - 한국과학기술원 : 생물공학과, 1984.2, [ [vi], 49 p. ]

URI
http://hdl.handle.net/10203/28161
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=64041&flag=dissertation
Appears in Collection
BS-Theses_Master(석사논문)
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