TY - JOUR
T1 - Development of shuttle vectors for evaluation of essential regulator regulated gene expression in Staphylococcus aureus
AU - Yan, Meiying
AU - Yu, Chuanxin
AU - Yang, Junshu
AU - Ji, Yinduo
N1 - Funding Information:
This work was supported by Grant AI057451 from the National Institute of Allergy and Infectious Disease.
PY - 2009/5
Y1 - 2009/5
N2 - We describe the construction of a series of shuttle vectors for Staphylococcus aureus. In order to determine transcriptional regulation by essential regulators, we constructed promoterless luxABCDE reporter system using a TetR-regulated antisense RNA expression vector, pJYJ909, which is composed of S. aureus plasmid pE194, the Gram- plasmid pUC18, a TetR regulatory cassette, and Pxyl/teto-driven yhcS antisense expression construct. The reformed shuttle vector was utilized to construct an opuCA promoter-luxABCDE fusion and simultaneously examine transcriptional regulation by measuring bioluminescence intensity during down-regulating yhcSR. In addition, we utilized the same plasmid, pJYJ909, and constructed a Pspac-driven constant expression system, which allows us to determine the complementary effect of overexpression of opuCA operon modulated by yhcSR. These plasmids provide important tools for elucidating regulatory mechanisms for genes that are essential for bacterial growth in S. aureus.
AB - We describe the construction of a series of shuttle vectors for Staphylococcus aureus. In order to determine transcriptional regulation by essential regulators, we constructed promoterless luxABCDE reporter system using a TetR-regulated antisense RNA expression vector, pJYJ909, which is composed of S. aureus plasmid pE194, the Gram- plasmid pUC18, a TetR regulatory cassette, and Pxyl/teto-driven yhcS antisense expression construct. The reformed shuttle vector was utilized to construct an opuCA promoter-luxABCDE fusion and simultaneously examine transcriptional regulation by measuring bioluminescence intensity during down-regulating yhcSR. In addition, we utilized the same plasmid, pJYJ909, and constructed a Pspac-driven constant expression system, which allows us to determine the complementary effect of overexpression of opuCA operon modulated by yhcSR. These plasmids provide important tools for elucidating regulatory mechanisms for genes that are essential for bacterial growth in S. aureus.
KW - Antisense RNA
KW - Promoter-reporter fusion
KW - Staphylococcus aureus
KW - TetR-regulated promoter
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U2 - 10.1016/j.plasmid.2009.02.001
DO - 10.1016/j.plasmid.2009.02.001
M3 - Article
C2 - 19245820
AN - SCOPUS:64749105627
SN - 0147-619X
VL - 61
SP - 188
EP - 192
JO - Plasmid
JF - Plasmid
IS - 3
ER -