Genome research of gut bacteria, how to analyze and how to apply

Abstract

After  development  of  new  generation  DNA  sequencers,  the  analyses  of  bacterial genome have been easy and familiar to microbiologist.  However, to utilize  vast amounts of data, it need  bioinformatics  and  reverse-genetics,  including  transformation  of  plasmid  DNA  into  target bacterium  and  gene  knockout  technique  using  homologous  recombination.  The  authors  proposed novel  transformation  technique  to  control  restriction  enzyme  reaction  using  plasmid  artificial modification  (PAM).  For  the  knockout  gene,  an  error  prone  PCR  has  been  employed  to  construct temperature sensitive plasmid.  Using this plasmid the knockout technique become extremely easier, than conventional method.