Enzymatic Synthesis of Peptidoglycan in Methicillin-Resistant Staphylococcus Aureus and Its Inhibition by Beta-Lactams
Total Page:16
File Type:pdf, Size:1020Kb
Enzymatic Synthesis of Peptidoglycan in Methicillin-Resistant Staphylococcus Aureus and Its Inhibition by Beta-lactams The Harvard community has made this article openly available. Please share how this access benefits you. Your story matters Citation Srisuknimit, Veerasak. 2019. Enzymatic Synthesis of Peptidoglycan in Methicillin-Resistant Staphylococcus Aureus and Its Inhibition by Beta-lactams. Doctoral dissertation, Harvard University, Graduate School of Arts & Sciences. Citable link http://nrs.harvard.edu/urn-3:HUL.InstRepos:42029540 Terms of Use This article was downloaded from Harvard University’s DASH repository, and is made available under the terms and conditions applicable to Other Posted Material, as set forth at http:// nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of- use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`WZ=YU\a* ZY* G(4B=-*EJF* ?$<()#&"&7&"()*(0*A$A&",(E1F<')*B"(#F)&3$#"#*BF*/H/L* Z]* G(4B=-*EJK* /H/L*'<&"-$*#$%")$*%$#",7$*")*&3$*&%')#A$A&",'#$*,(8'")*"#* ZZ* )$<$##'%F*0(%*A$A&",(E1F<')*<%(##1")5")EG* G(4B=-*EJL* /7%"0"<'&"()*(0*!/,#*-.*+*/H/L'*`LX=\\]a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b;6*$Q&%'<&$,*"()*<3%(8'&(E%'8*(0*!/,#*-.*+*/H/L*2X[\+** UXL* +B225J*G(4J*K* +*,"%$<&*&%')#A$A&",'#$*'<&"-"&F*'##'F*$)'B1$#*<3'%'<&$%"9'&"()*(0* UXV* ")3"B"&(%F*A(&$)<"$#*(0*,"00$%$)&*B$&'=1'<&'8#* +B225J*G(4J*L* J"8$=<(7%#$*')'1F#"#*(0*/H/L*%$'<&"()*I"&3*T1FW=O"A",*PP*")* UXX* JT'#$*B700$%* * * * * * * * * * * * * * * * * D(.'*/0*>&:5-.*