Inactivation Gene waaF Aa using Neighbour Joinning Tehnique

Objectives:LPS is an essential virulent determinant expressed by Gram negative bacteria, including Aggregatibacter actinomycetemcommitans(Aa) periopathogenic bacteria.The aim of this study was to inactivated Acyl Transferase Gene(waaF)which are involved in the biosynthesis of the inner core of the bacteria LPS.Method:To obtain the waaF mutant The Neighbour Joinning Method was used to construct a suicide vector,which carries a kanamicyn resistance gene inserted into the bacterial DNA fragment encoding the waaF gene.Result: This study found that the neighbour joinning method could succesfully be used to construct a suicide mutant construct.The resulted suicide vector were further used to naturally transform Aa isolated from oral sample,which after homologous recombination resulted in a kanamicyn resistant mutant.Conclusion:It can be recommended the neighbour joinning method could be used as a suitable method in activating any coding region located in Aa genome.