1. Two Newly Discovered genes may encode for proteins associated with the Proteasome - Neville Campbell
Intro. Good evening everyone, my name is Neville Campbell and for the next 8 or so minutes I will be talking to you about two newly discovered genes that may encode for proteins associated with the proteasome.

2. Human Genome Project
Identify all the approximately 30,000 genes in human DNA.
Determine the sequences of the 3 billion chemical base pairs that make up human DNA
New studies in Genomics, Proteomics and Bioinformatics
The human Genome project began in 1990 and was completed thirteen years later. The project had a number of objectives, two of which were to identify all the genes in the human DNA and to determine the sequences of the chemical base pairs that makeup the human DNA.
This undertaking has led to a an explosion of studies and research in the areas of genomics, proteomics and bioinforamtics. The paramount thrust here is to understand and address many of the mysteries that surround the diseases which we are faced with today.

3. CGI – 112 & NOC4 Neighbor of Cytochrome oxidase 4 (NOC4)
Comparative Gene Identification (CGI -112)
Currently Dr Bachman and few students including myself are investigating two genes, NOC4, which is located next to the cytochrome oxidase IV subunits on chromosome 14 and Comparitive Gene Identification 112 which was discovered using bio-informatics. Both genes encode for proteins that are approximately 40% identical in amino acid sequence. Because of this similarity, we believe that they evolved from a common ancestral gene and may be apart of the same family.
It is our hope to determine possible functions of the proteins that these genes encode for, however such a task is lacking in simplicity because of the complex nature of genes and the processes that they impact.
This task of identifying function however is marked with trail and error, hypothesis rejection, and frustration, but its benefits are of tremendous importance. Especially, if the researcher determines a correlation between the gene understudy and a disease or health problem. A combination of techniques are used in accessing gene function, one of these is the use to Green Fluorescent fusion proteins.

4. Green Fluorescent Protein (GFP)
Derived from a jellyfish
Ability to fluoresce green when exposed to light of certain wave lengths
GFP is derived from the bioluminescent jellyfish Aequorea victoria. This jelly fish is able to fluoresce green light when it is exposed to light of certain wave lengths. This phenomenon is due to a protein present in the creatures structure that has the ability to transition back and forth between different energy levels. This protein has provided researcher with a powerful tool with which to study sub-cellular localization of proteins. The localization of proteins is important because they indicate a possible function of the that protein. For example if a protein localizes in the mitochondria then one can say that this protein has to do with the various processes in the mitochondria. Library Example.

5. Method – how is GFP used as a reporter?
HeLa cells
Transfection
Microscopy
Pre-Transfection HeLa cells
Post-Transfection
The green fluorescent protein is fused to the gene of interest though genetic engineering. This construct is then introduced to Hela cells. These cells are cervical cancer cells that are derived from a woman named Henrietta Lacks, who died in The have had remarkable success in culturing so they are wide used in research today. They have to be grown under special conditions in the lab for a few weeks prior to experimenting. When cells are ready we introduce the GFP construct tot the cells through a process called transfection. When the transfection process is complete the cells are fixed then placed on glass slide to be view under a fluorescent microscope. Photographs of the localization pattern are then taken. Here is a look at some of the localization patters.

6. Localization patterns of NOC4 & CGI -112
We have noticed similarity between our images and those observed from immunofluoresce images of proteasome localization we
CG1-112

7. Immunofluorescence image of proteasome
Based on the localization patterns we believe that these protein are related to the proteasome in structure of in function.

8. What is a proteasome Protein Degradative complexes Different Subunits
Proteasomes are protein degradative complexes that responsible for protein turn over and in some cases antibody production in cells. They are made up of different sub units that have different functions. The major component of the proteasome is the 20s core which is featured in figure one. The aspect is responsible for protein degradation. The other tow aspects of the proteasome are the 11s and the 19s subunits. The 11s subunit is involved in antigen production and the 19s subunit is involved with the unbiquitination process. We believe the our proteins are with being broken down by the proteasome or the become apart of the proteasome structure.

9. Future Experiments