Construction and Analysis of a Modified Tn4001Conferring Chloramphenicol Resistance inMycoplasma pneumoniae

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• 作者：Hahn ; Tae-wook ; Mothershed ; Elizabeth A. ; Waldo III ; Robert H. ; Krause ; Duncan C.
• 刊名：Plasmid
• 出版年：1999
• 出版时间：March, 1999
• 年：1999
• 卷：41
• 期：2
• 页码：120-124
• 全文大小：53.0 K

文摘

TheStaphylococcus aureustransposon Tn4001and derivatives thereof have been transformed successfully in several mycoplasma species. In order to expand the versatility of Tn4001for other genetic manipulations and for use in mycoplasma species resistant to gentamicin (Gm), chloramphenicol acetyltransferase (Cat) fromS. aureuswas evaluated as a selectable marker. Thecatgene was cloned in both orientations into a modified Tn4001and transformed intoMycoplasma pneumoniae,conferring resistance to Cm and Gm. Replacement of the gene for Gm^Rin Tn4001withcatlikewise conferred Cm^Rwhen transformed intoM. pneumoniae.The minimum inhibitory concentration to Cm in transformants withcatderivatives of Tn4001was 300–500 μg/ml, and Cat enzyme activity was demonstrated by using a fluorescent substrate.