Well, the time has come to say goodbye.  First I want to answer a few of the questions that are asked regarding this internship opportunity:

Was this a good experience?  Absolutely   

Would I do this again?  Positively                                                      

Did I learn anything?  More than I ever thought I would.            

Where the people good to work with and for?  Words can't express how much I appreciate my mentor and the entire Lab!!!

Would I recommend an internship to other students?  Yes, take the opportunity and run with it.  Be active, involved and eager to learn, you will be amazed at what can happen.                                                 

Dr.H, Dr.C, Vic, Jack and "cats"……………..Thank you!!!!!!                                                                                                                               

For everyone else, let's review what we have learned:

Most people have it.

Immunity doesn't exist.

CMV (herpes, the gift that keeps on giving)

Really serious.

Only a few treatments, NO CURE

Birth defects

Illness

Our lab studies

Lots of cool stuff

Opportunities galore

Green glow is our favorite color

Y because science is cool!!!!

This week I took the membrane that contained the protein results from the GFPSCP experiment and tested it for the presence of another protein that is present in virions. If this protein is detected in the same area as my GFPSCP that will be a good indicator that the construct is working the way we hoped. If the protein is detected passed the areas where the GFPSCP was detected, that would imply that the construct is not being incorporated into the virion and is therefore not working.

The good news is that the experiment worked and I was able to detect the additional protein.

The bad news is that the protein was detected everywhere!!!!!  Before GSCP, after GSCP, everywhere. The results are so strong that I am not able to draw any conclusions from these results.

Oh well!  Back to the old drawing board.

My time in the spot light as a project presenting mini scientist has almost come to an end. This week I was back to Lisa the average everyday dish washing lab tech. I am so incredibly lucky and blessed to work with this group of scientists that I am truly content at this point just to wash their dishes and mix their chemicals.

I will continue to work on my project in the lab.  My time as a blogging intern will soon come to an end. The school starts thanksgiving break tomorrow and when we return it will be finals and my last blog should be around Dec. 4^th.

I will be in the lab this week and will hopefully have more GFPSCP news next week.  Have a great holiday!!!!!!!

Believe it or not, Dr. H showed up in Richmond at 4:00pm and slipped me a GFP picture during the meeting!!!!!!  Thank you Lord!!!! 

Note to Everyone:

A good scout is always prepared.  I had brought my portable paper cutter, the spray adhesive and an addition board to mount the slide on.

At the 4:20 break I ran outside, cut, sprayed and mounted the picture.

Taa Daa!!  The poster was up at 4:30.  Just in time for the butterflies in my stomach to take flight. 

From 6:30 till a little after 8pm I stood by my poster waiting to explain my research and answer questions.

I am very excited about how things went. 

I had a good steady flow of traffic at my poster (It's very discouraging to be standing there alone with no one looking at your work).

This was a great experience.  You get input on your work, suggestions, ideas and compliments.  The VA ASM meeting is a very student friendly environment.  The meeting is an opportunity to show case your work, network in the research field and you get a chance to see other research.  The professors and scientists attending the meeting see this as an opportunity to share ideas, network and to look for potential Grad students or employees.  Everyone really benefits.

I am very thankful for this opportunity.

It is Friday morning.

We are leaving at 9:30am for Richmond.

The meeting starts at 1:00pm.

The vendor has NOT shown up to fix the confocal.

Dr. H has decided to stay back for a seminar at EVMS and hopefully give the vendor time to show up.

She is going to try to get a picture and bring it later.

Friday afternoon traffic.

Interstate 64

My poster has to set up by 4:30.

No I'm not stressing.  NOT AT ALL…….

The American Society for Microbiology Meeting (ASM) is very close.  I have been working on putting my poster presentation together.  I am a little worried, I still need a confocal microscope picture of my cells that were transfected with GFPSCP.  I know your probably thinking so get it done, easier said that done:

1.      Confocal Microscopy is very expensive and special training is required (Lab techs don't have access till they grow up to be bigger scientists)  Thank Heavens I have Dr. H to help me.

2.      Dr. H can't help me yet because the mercury lamp on the microscope has burned out and we are waiting for the vendor to fix it.  (SOOOOOO typically my luck, just to keep things interesting)

I am preparing two versions of the poster, one with GFP pictures and one without.  I just want to be prepared either way.

GFPSCP is glowing green!!!!!  I am now running tests to determine where the green glow is coming from.   (No photos yet, sorry) This part of the process has taken longer than I hoped.  I may have results by the end of this week.      

Now for the interesting news, today I will begin putting all of the results from this project into a poster presentation. I have the unique opportunity to present my research to the Virginia Chapter of the American Society of Microbiology. I am going to be doing an Undergraduate poster presentation.  I will also be a nervous wreck.  During the presentation I will stand beside my poster ready to explain the project and hopefully be able to answer questions from Scientists, Professors and other students at the meeting. Did I mention nerves, stage fright? I think I feel a case of agoraphobia coming on.   

OK that may be a little dramatic, I feel like I know just enough to be dangerous.  The truth is, I know I will be OK. Dr. H and two of the PhD students will be making oral presentations and will be very much available to help me answer questions.  This best part of doing this experiment has been the encouragement and support of the people I work with.

Sorry for the delay in updates.  One of the scientists here at work passed away last week.  Our lab planned and provided his memorial service and reception so we didn't get much research done.  I will be running test on GSCP tuesday and should be updating the blog this weekend.  Thanks for your patience.

Guess what?  GFPSCP is finally starting to act right….. we are not going to discuss the extent of operator error involved with slowing down the progress, its just nice that to know that I work for very patient scientists.                                                                                                                                                                                                                                                                        

I think this would be a good time to talk about why I am doing this project.                                                                                                                                                                                                                           

As I explained at the beginning of this project, we work on cytomegalovirus (CMV), a herpes virus.  Our lab has already established that if you remove one particular gene from the viral DNA the virus does not grow well in certain cells.

                                                                                                                                                                                                                                                                                                                                                                                                                           The pictures below show cells infected with CMV, the black spots with a circle around it are single virions. The cell on the left was infected with the mutant virus (missing a single gene).  The cell on the right was infected with the "normal" wild type virus.

                                                                                                                                                                      As you can see the mutant virus resulted in ONE virion versus several in the cell infected with wild type virus.  Fewer virions means fewer capsids.

This is a closer look at a herpes virion.

The large circle in the middle is the capsid, a structured protein shell around the DNA.

                                                                                                                                                                                                                      We believe that the mutant virus results in unstable capsids.  I am tagging the small capsid protein gene as a tool to follow the capsid during wild type and mutant infections.  If the GFPSCP construct works as expected we will actually be able to see the capsid glow green. Hopefully I will have GFP (Green glowing) pictures for you next week.

Finally! I am ready to share some of my photos.                                                   

First let's review:

These photos are of cells that contain proteins produced by the flag tagged small capsid protein gene. The cells were treated with antibodies that cause the FSCP protein to appear red.                                                                                                                           

The two photos below are of normal cells that are not infected with MCMV. 

I hope that you can see that the SCP protein is all through out the cell with few concentrated areas.                              

We are creating these constructs in an effort to study how scp works inside infected cells.  The following cells were infected with MCMV and treated with two different antibodies:

• one to cause SCP to appear red
• one that causes E1 proteins to appear green.

We know how proteins produced by E1 participate in viral replication. I used E1 as a marker to see if the SCP proteins appear in the same areas.

        When the cells are infected, the SCP is under the control of the virus and responds in a more natural manner.                      

        We found cells showing concentrated areas of SCP that appear to surround the E1 proteins. The first picture is SCP, the second is E1 and the third is a merge of the first two.

We also found cells with slightly yellow areas, these are areas of colocalization where the two proteins are in the same space at the same time.   

                                                                                                                                                                                                                                                                 
I will explain all of this soon.  I just wanted to share some of my results.

Be back soon with info on the GFPSCP.

Hurray for FSCP!!  So far so good