TY - JOUR
T1 - Gene essentiality testing in mycobacterium smegmatis using specialized transduction.
AU - Bhatt, Apoorva
AU - Jacobs, WR
PY - 2009/1/1
Y1 - 2009/1/1
N2 - Conditional expression-specialized transduction essentiality test (CESTET) is a genetic tool used to determine essentiality of individual genes in Mycobacterium smegmatis. CESTET combines specialized transduction, a highly efficient gene knockout method, with the utility of the inducible acetamidase promoter. In a merodiploid strain containing a second integrated copy of an essential gene under the control of the acetamidase promoter, transductants (gene knockouts of the native chromosomal copy of a gene) are obtained only in the presence of acetamide in the selection medium. Furthermore, effects of loss of essential gene function can then be studied by growing the transductants in medium depleted of acetamide.
AB - Conditional expression-specialized transduction essentiality test (CESTET) is a genetic tool used to determine essentiality of individual genes in Mycobacterium smegmatis. CESTET combines specialized transduction, a highly efficient gene knockout method, with the utility of the inducible acetamidase promoter. In a merodiploid strain containing a second integrated copy of an essential gene under the control of the acetamidase promoter, transductants (gene knockouts of the native chromosomal copy of a gene) are obtained only in the presence of acetamide in the selection medium. Furthermore, effects of loss of essential gene function can then be studied by growing the transductants in medium depleted of acetamide.
U2 - 10.1007/978-1-59745-207-6_22
DO - 10.1007/978-1-59745-207-6_22
M3 - Article
C2 - 20560066
SN - 1940-6029
VL - 465
SP - 325
EP - 336
JO - Methods in molecular biology
JF - Methods in molecular biology
ER -