Effect of DnaA protein on lacZ fusion and Characterization of nrdAB Promoter

Narasimha Rao Uda & Rajashekhar Kamalampeta

Studenteropgave: Semesterprojekt

Abstrakt

ABSTRACT DnaA is known to be the most important protein involved in the initiation of DNA replication in bacteria. It is also found to be involved in transcription regulation. It is well known that DnaA binds to the DnaA boxes. Some experiments have shown that the mutation in DnaA boxes brings down the expression of the genes, indicating that DnaA has some role to play in this. In this experiment we put an effort to show the effect of DnaA on expression of a gene, using lacZ fusions. lacz fusion and plasmid carrying dnaA are introduced into a strain to study the effect of DnaA along with a strain with no dnaA for comparision. Increasing concentrations of IPTG is used to induce these strains and are analyzed with PAGE and Immuno blot. β- galactosidase assays are used to see the expression. Also carried the experiments to characterize the nrdAB promoter. Strains (with DnaA and lacz) carrying various regions of the nrdAB promoter in the plasmids are designed and are grown exponentially. The samples are analyzed using β- galactosidaseassay, GFP and with flowcytometer. Some positive results are obtained with GFP analysis.

UddannelserBiologi mhp. gymnasiet, (Bachelor/kandidatuddannelse) Bachelor el. kandidat
SprogEngelsk
Udgivelsesdato1 jun. 2006
VejledereTove Atlung

Emneord

  • Molecular Biology