Abstract
A new shuttle vector, pRH3 (8.7 kb), was constructed for use in Prevotella/Bacteroides host strains. This vector combines the pRR12 replicon from P. ruminicola, pBluescript sequences and a tetQ marker gene for selection in Prevotella/Bacteroides hosts. Following insertion of a fragment carrying an endoglucanase/xylanase gene from P. ruminicola 23 into the multiple cloning site, the resulting construct, pRH3X, was introduced into B. vulgatus 1447, B. uniformis 1100 and P. ruminicola 2202. This resulted in increases of between 4 and 50-fold in CM-cellulase and xylanase activities in cells grown with glucose. In contrast activities were barely detectable for the same construct in E. coli DH5 alpha. Most of the total xylanase activity produced was found within the cell in P. ruminicola 2202 and B. vulgatus 1447 transformed with pRH3X, and in P. ruminicola 23, An osmotic shock experiment indicated that a significant proportion of the xylanase activity in B. vulgatus 1447 cells carrying pRH3X was periplasmic.
Original language | English |
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Pages (from-to) | 417-424 |
Number of pages | 8 |
Journal | Journal of Applied Bacteriology |
Volume | 79 |
Issue number | 4 |
Publication status | Published - Oct 1995 |
Keywords
- TETRACYCLINE RESISTANCE
- ESCHERICHIA-COLI
- SHUTTLE VECTOR
- DNA
- BACTERIA
- RUMEN
- MANIPULATION
- SEQUENCE
- STRAINS
Cite this
EXPRESSION OF A CLONED CELLULASE XYLANASE GENE FROM PREVOTELLA-RUMINICOLA IN BACTEROIDES-VULGATUS, BACTEROIDES UNIFORMIS AND PREVOTELLA-RUMINICOLA. / DANIEL, A S ; MARTIN, J ; VANAT, I ; WHITEHEAD, T R ; FLINT, H J .
In: Journal of Applied Bacteriology, Vol. 79, No. 4, 10.1995, p. 417-424.Research output: Contribution to journal › Article
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TY - JOUR
T1 - EXPRESSION OF A CLONED CELLULASE XYLANASE GENE FROM PREVOTELLA-RUMINICOLA IN BACTEROIDES-VULGATUS, BACTEROIDES UNIFORMIS AND PREVOTELLA-RUMINICOLA
AU - DANIEL, A S
AU - MARTIN, J
AU - VANAT, I
AU - WHITEHEAD, T R
AU - FLINT, H J
PY - 1995/10
Y1 - 1995/10
N2 - A new shuttle vector, pRH3 (8.7 kb), was constructed for use in Prevotella/Bacteroides host strains. This vector combines the pRR12 replicon from P. ruminicola, pBluescript sequences and a tetQ marker gene for selection in Prevotella/Bacteroides hosts. Following insertion of a fragment carrying an endoglucanase/xylanase gene from P. ruminicola 23 into the multiple cloning site, the resulting construct, pRH3X, was introduced into B. vulgatus 1447, B. uniformis 1100 and P. ruminicola 2202. This resulted in increases of between 4 and 50-fold in CM-cellulase and xylanase activities in cells grown with glucose. In contrast activities were barely detectable for the same construct in E. coli DH5 alpha. Most of the total xylanase activity produced was found within the cell in P. ruminicola 2202 and B. vulgatus 1447 transformed with pRH3X, and in P. ruminicola 23, An osmotic shock experiment indicated that a significant proportion of the xylanase activity in B. vulgatus 1447 cells carrying pRH3X was periplasmic.
AB - A new shuttle vector, pRH3 (8.7 kb), was constructed for use in Prevotella/Bacteroides host strains. This vector combines the pRR12 replicon from P. ruminicola, pBluescript sequences and a tetQ marker gene for selection in Prevotella/Bacteroides hosts. Following insertion of a fragment carrying an endoglucanase/xylanase gene from P. ruminicola 23 into the multiple cloning site, the resulting construct, pRH3X, was introduced into B. vulgatus 1447, B. uniformis 1100 and P. ruminicola 2202. This resulted in increases of between 4 and 50-fold in CM-cellulase and xylanase activities in cells grown with glucose. In contrast activities were barely detectable for the same construct in E. coli DH5 alpha. Most of the total xylanase activity produced was found within the cell in P. ruminicola 2202 and B. vulgatus 1447 transformed with pRH3X, and in P. ruminicola 23, An osmotic shock experiment indicated that a significant proportion of the xylanase activity in B. vulgatus 1447 cells carrying pRH3X was periplasmic.
KW - TETRACYCLINE RESISTANCE
KW - ESCHERICHIA-COLI
KW - SHUTTLE VECTOR
KW - DNA
KW - BACTERIA
KW - RUMEN
KW - MANIPULATION
KW - SEQUENCE
KW - STRAINS
M3 - Article
VL - 79
SP - 417
EP - 424
JO - Journal of Applied Bacteriology
JF - Journal of Applied Bacteriology
SN - 0021-8847
IS - 4
ER -