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동의어 포함
A biocontrol bacterium Bacillus licheniformis N1 grown
in nutrient broth showed no chitinolytic activity, while
its genome contains a gene which encodes a chitinase.
The gene for chitinase from B. licheniformis N1 was amplified
by PCR and the deduced amino acid sequence
analysis revealed that the chitinase exhibited over 95%
identity with chitinases from other B. licheniformis
strains. Escherichia coli cells carrying the recombinant
plasmid displayed chitinase activity as revealed by the
formation of a clear zone on chitin containing media,
indicating that the gene could be expressed in E. coli
cells. Chitinase gene expression in B. licheniformis N1
was not detected by RT-PCR analysis. The protein was
over-expressed in E. coli BL21 (DE3) as a glutathione Stransferase
fusion protein. The protein could also be
produced in B. subtilis 168 strain carrying the chitinase
gene of N1 strain. The crude protein extract from E. coli
BL21 carrying GST fusion protein or culture supernatant
of B. subtilis carrying the chitinase gene exhibited enzyme
activity by hydrolyzing chitin analogs, 4-methylumbelliferyl-
β-D-N,N'-diacetylchitobioside and 4-methylumbelliferyl-
β-D-N,N',N''-triacetylchitotrioside. These results
indicated that even though the chitinase gene is not expressed
in the N1 strain, the coding region is functional
and encodes an active chitinase enzyme. Furthermore,
B. subtilis 168 transformants expressing the chitinase
gene exhibited antifungal activity against Fulvia fulva
by suppressing spore germination. Our results suggest
that the proper engineering of the expression of the
indigenous chitinase gene, which will lead to its expression
in the biocontrol strain B. licheniformis N1, may
further enhance its biocontrol activity.*표시는 필수 입력사항입니다.
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도서위치안내: 정기간행물실(524호) / 서가번호: 국내18
2021년 이전 정기간행물은 온라인 신청(원문 구축 자료는 원문 이용)
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