PERUBAHAN SEBAGIAN NUKLEOTIDA GEN β-LAKATAMASE PADA Escherichia coli RESISTEN AMOKSISILIN-KLAVULANAT SECARA IN VITRO
In Indonesia there are many E. coli resistant amoxicillin-clavulanate exist, and the fundamental causes are the hyperproduction of TEM-1 β-lactamase enzyme and or IRT (Inhibitor Resistance TEM). This research is aimed to observe the alteration of nucleotide gene order which causes the alteration of β-lactamase amino acid enzyme order in the fourth isolates clinical E. coli sensitive to amoxicillin-clavulanate. Plasmid that success isolated from the fourth E. coli amox-clavS and four E. coli amox-clavR are used as the template for the amplification of β-lactamase gene through PCR technique. Nucleotide sequences directly determined by using automatic DNA-sequencer.As the result it is found out that all nucleotide in all gene very homology to TEM-1 β-lactamase gene. The comparation analysis of nucleotide gene sequence from sensitive and resistance of amoxicillin-clavulanate shows inform of nucleotide difference over 1 until 3. Two of them located in regulation area and down stream of early translation and also the area of enzyme code, and the resistance happens because the mutation influences gene expression level. One of them located on the area enzyme code and the resistance happens because of the very often use of amino acid codon. While the other do not have any alteration in its nucleotide gene and the resistance that is happens may be caused by the mutation in other area, such as porin gene or amplification gene. All nucleotide alteration in the area of enzyme code does not cause any alteration on amino acid enzyme sequences.In conclusion : The effect of the exposure of amoxicillin-clavulanate in vitro can cause alteration E. coli from sensitive to resistant amoxicillin-clavulanate, and the alteration is caused by the alteration of nucleotide sequences but this does not cause any alteration in β-lactamase amino acid enzyme sequences..
Keywords: E. coli, Resistant amoxicillin-clavulanat, Nucleotide β-lactamase gene sequences
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