Analysis And Validate The Replication Origin Of Burkholderia Multivorans ATCC17616

The replication of DNA is a copy process which happens before the cell divisionof double-stranded DNA，is the basic of the biological genetics. The replication ofDNA including three steps that are priming, extending and termination. The origin ofreplication is a central event of the life cycle of the bacterial cells. So theidentification of the oriC is such an important foundation stage for the research of theprocess of DNA replication. At present, the study on the determine and analysis theoriC of bacterial has not been widely carried out. This work is basic on the Ori-Finderwhich was developed by TUBIC. Analysis the oriC of Burkholderia multivoransATCC17616and then verify the accuracy of the prediction results through theinteraction of protein-DNA. This will lay the groundwork for the widely used in theprediction of the bacterial genome oriC.First, construction of ATCC17616oriC cloning vector named pSURE-T-oriCaccording the informations from Ori-Finder, using the methods of molecular cloning.Second, construction of ATCC17616dnaA expression vector named pET28a-dnaAaccording the information from NCBI. Last, verify the interaction of dnaA and oriCusing EMSA. From the result, we can find that there was positive result between dnaAand oriC from the same chromosome. Two other oriCs from the remaining twochromosomes may have interaction with another proteins, more experimentalrequired.