Insertion of an additional origin in E. coli bacterial chromosomal DNA

Mads Joachim Hansen, Niclas Helge Ulla Nilsson, Jacob Tofte & Laura Emilie Bjergbæk Olsson

Studenteropgave: Fagmodulprojekt

Abstrakt

A secondary origin site is not a common occurrence in E. coli bacteria and therefore it is not a well-studied field. We intend to supply an E. coli strain with a plasmid containing an additional origin of replication in a transposon in order to transpose the origin of replication into chromoso-mal DNA of an E. coli strain. This is achieved by ligating a fragment containing the origin of repli-cation into a pBT20 plasmid and then electroporate that into a recipient strain. This can then be used to sequence the entire genome of the E. coli with or without an additional origin site. Com-paring these two different libraries will give the opportunity to see where in the chromosomal DNA the origin site cannot be transposed without causing vital damage to the cell. This project provides evidence of a successful conjugation of the pBT20 plasmid both with and without the additional origin of replication. It has not been possible to provide sufficient evidence to confirm that a transposition has happened after conjugating and electroporating the respective vectors into a recipient cell though some results indicate that a transposition has taken place.

UddannelserMolekylærbiologi, (Bachelor/kandidatuddannelse) Bachelor
SprogEngelsk
Udgivelsesdato17 dec. 2019
Antal sider41
VejledereOle Skovgaard