植物保護─BACILLUS幾丁質分解酵素基因在PSEUDOMONAS根圈細菌之表現分析(2/3)..
Date Issued
2000
Date
2000
Author(s)
DOI
892317B002002
Abstract
We plan to transfer a Bacillus chiA
gene into Pseudomonas rhizobacteria for
the expression of chitinase and
tranlocation of chitinase to the outer
membrane of bacterial cells. We expect
the transformed Pseudomonas
rhizobacteria will express antifungal
activity or be with enhanced antifungal
activity. A DNA segment containing the
2
sequences for signal peptide and
transmembrane domain (ST sequence)
was amplified by PCR, cloned into
pGEM-T easy vector and fused with
chiA gene segment of different lengths.
The construct was subcloned into
pMMB6. The plasmid pMMST2000
containing full length of chiA gene with
ST sequence expressed chitinase in E.
coli Top10F’ as shown in glycol chitin-
SDS-PAGE, on trypan blue-glycol chitin
medium, and in fluorometric assay,
indicating that ST sequence would not
affect the expression of chitinase in
bacterial cells. Since E. coli
Top10F’(pMMST2000) had a good
growth activity and made clearing of
chitin medium just beneath the colonies,
we assumed that the ST sequence could
drive the chitinase to the membrane. In
the next, plasmid pMMST2000 will be
transferred into Pseudomonas
rhizobacteria to examine the antifungl
activity of transformants.
Subjects
chitinase
chitinase gene
rhizobacteria
genetic engineering
gene
expression vector
expression vector
biocontrol
Publisher
臺北市：國立臺灣大學植物病理與微生物學系暨研究所
Type
other
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