DNA Damage Investigated by Restriction Enzyme Cleavage
Poster Number
12
Format
Poster Presentation
Abstract/Artist Statement
Damage to the DNA contained in human chromosomes is one of the main causes of cancer. Examples of DNA damage are breaks in the chromosomes or chemical alterations of the bases, the letters of the genetic code. DNA damage can interfere with biological processes and the enzymes that catalyze them. Thus, enzymes can be used as a way of studying DNA damage. Restriction enzymes are proteins that cut DNA at specific sequences. If one of the code letters is blocked, they can not cut the DNA. Restriction enzymes produce a specific pattern of DNA fragments when used in conjunction with appropriate pieces of DNA. We used a readily available DNA structure called a plasmid for our studies. There are several advantages to using a plasmid. For example, it can be produced by bacteria in large quantities. We found the suitable restriction enzymes for cutting our plasmid by searching appropriate databases. Then we cut the plasmid with the enzymes and detected the restriction fragment by agarose gel electrophoresis and a fluorescent stain. We applied this method to the study of DNA damage caused by cis- platin, a drug used for killing cancer cells. The drug, cis-platin, usually reacts readily with DNA and does not require additional cellular factors to produce very high levels of DNA damage. We found that treating the plasmid with cis-platin prevented cutting with restriction enzymes. We extended our study to other DNA molecules that include fragments of genes that are known to be damaged in certain cancers. Next, we will study various conditions, including food that may increase or decrease the damage caused by cis-platin. Results from these studies may lead to recommendations for patients undergoing cis-platin chemotherapy.
Location
Pacific Geosciences Center
Start Date
30-4-2005 1:00 PM
End Date
30-4-2005 3:00 PM
DNA Damage Investigated by Restriction Enzyme Cleavage
Pacific Geosciences Center
Damage to the DNA contained in human chromosomes is one of the main causes of cancer. Examples of DNA damage are breaks in the chromosomes or chemical alterations of the bases, the letters of the genetic code. DNA damage can interfere with biological processes and the enzymes that catalyze them. Thus, enzymes can be used as a way of studying DNA damage. Restriction enzymes are proteins that cut DNA at specific sequences. If one of the code letters is blocked, they can not cut the DNA. Restriction enzymes produce a specific pattern of DNA fragments when used in conjunction with appropriate pieces of DNA. We used a readily available DNA structure called a plasmid for our studies. There are several advantages to using a plasmid. For example, it can be produced by bacteria in large quantities. We found the suitable restriction enzymes for cutting our plasmid by searching appropriate databases. Then we cut the plasmid with the enzymes and detected the restriction fragment by agarose gel electrophoresis and a fluorescent stain. We applied this method to the study of DNA damage caused by cis- platin, a drug used for killing cancer cells. The drug, cis-platin, usually reacts readily with DNA and does not require additional cellular factors to produce very high levels of DNA damage. We found that treating the plasmid with cis-platin prevented cutting with restriction enzymes. We extended our study to other DNA molecules that include fragments of genes that are known to be damaged in certain cancers. Next, we will study various conditions, including food that may increase or decrease the damage caused by cis-platin. Results from these studies may lead to recommendations for patients undergoing cis-platin chemotherapy.