Enterobacter cloacae subsp. cloacae (Jordan) Hormaeche and Edwards, subsp. nov.
Depositors
KJ Shaw
Applications
The 400bp ClaI/EcoRV fragment is recommended as a hybridization probe for aac(3)-VIa.
Vector
Construct size (kb): 5.05
Insert
DNA: genomic
Insert lengths(kb): 2.077
Gene product: acetyltransferase [aac(3)-VIa]
Insert Size (kb)
2.077
Media
ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Biosafety Level
1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Comments
Restriction digests of the clone give the following sizes (kb): ClaI/EcoRV--3.6, 1.2, 0.4; EcoRV--4.1, 1.1; ClaI--3.6, 1.6.
The insert contains the following restriction sites (approximate kb from the 5' end): SspI--0.41; ClaI--0.60; BglI--0.96, 1.75; EcoRV--1.00; XhoI--1.03, 1.79; SacII--1.72.
Two rounds of gel purification of the restriction digest generated probe is necessary. Failure to do so often results in high backgrounds (due to flanking regions in the plasmid insert) and false positives with clinical E. coli strains.
The 400bp ClaI/EcoRV fragment is recommended as a hybridization probe for aac(3)-VIa.
References
Shaw KJ, et alThe application of molecular techniques for the study of aminoglycoside resistanceIn: Shaw KJ, et alMethods in molecular medicine: molecular approaches for the diagnosis and investigation of bacterial diseasesTotowa, NJHumana Presssubmitted, 1996
