07/06/2007

Today was exciting. Our x-ray diffraction pattern was used on a cryo lysozyme crystal and input into HKL 2000. This program was used to get a 3- D data set of the crystal. With the information, we created an electron dot configuration and further created our "protein picture" from work we had all done. Additionally, all our crystals have been doing well. The researchers are all relatively impressed with us and willing to share a multitude of techniques. Next week, we will be working with PCR, and Western blot analysis and learning more about DNA shuffling so we can share our FIRST-HAND knowledge with our students regarding these currciulum topics. I know that when I convey learning to my students based upon my personal field experience, they always do well because I have examples, activities and enthusiasm for the subject. I only wish this could go on longer since there is so much I would like to learn!

7/5/07

We checked for phycocyanin crystals. All wells had great bluish crystals that resembled hairs or nails depending upon your perspective. We decided to wait before putting those through the x-ray machine in the hopes that they would grow even larger. As such, we used our lysozyme crystals for x-ray diffration and watched the x-ray machine work as observers since that is a more dangerous piece of equipment. The first crystal gave a relatively poor diffraction pattern despite its look on the outside. I guess- in crystals- as well as in people- it's whats in the inside that counts. We then waited for the next pattern and while we did that, we took more pictures of these new crystals using the microscope camera once again, only this time we took pictures of all wells. Tomorrow, we will be diffracting the crystals under cryo conditions- i.e. liquid nitrogen or propane- in the hopes that this will help the crystals retain more moisture. However, too much humidity is also a problem so it is like Goldilock's theorem where you have to get it "just right." My question is this: how do you get this beautiful colorful picture of the structure of these membrane proteins showing the symmetry and ribbon-like structure of the amino acids? That is the question that I will ask next and possibly do some independent research upon myself as well.

7/3/07

Today, we did vapor diffusion technique with the real thing in the hopes of making large crystals for x-ray diffraction. We also used the spectrometer to determine the purity of the proteins being used based upon different dilutions. More importantly, we got to chatting about our own personal experiences. I am starting to feel more comfortable and confident with this process.

7/2/07

Lots of lecture today and I was very excited to find out the implications that this research has. With the splitting of water comes the vast possibility of endless energy and power for our future growing population. Humans are so cool. I look at this from an ecological perspective indeed which is why humans have not yet reached carrying capacity and I discuss this with my students as part of our standards all the time. Additionally, this has implications for drug interactions since humans are composed of 60% membrane proteins and drugs target these in particular. Plants are even more complicated- again another twist on the order and complexity of our classification system. It would indeed indicate that plants are much more complex and have to be especially in their defense mechanisms since they cannot move.

Additionally, we worked an ultra-centrifuge today and will be working with x-ray diffraction soon. The patterns are truly beautiful... hauntingly so- since they are so complex. Revisting vocabulary from my MS. Am very excited about the ecological standpoint of this!

Crystals are beautiful and my students would love to create them and I could then extrapolate upon solubility curves, titration, salt concentrations etc.

Techniques

Vapor diffusion technique- nice colorful crystals, batch technique- lots of crystals, and interface technique- more difficult to use with High School?

7/1/07

Just trying to add pix to my blog and the wiki. Having trouble with that since they often require a link or a URL and am not used to doing that. Will consult with my group tomorrow.

6/29/07

Found crystals usable for x-rays at A3, A4, D3 from vapor crystallation technique. Will be making photos of the crystals. Sat in on grad seminar. Dr. Fromme served cake to grad students for their birthday. The atmoshere was one of comraderie and collaboration. Student Rob Lawrence was having some trouble with his data- overexpressing a C subunit of spinach in E. coli. Several suggestions were given and an effort was sincerely made to explain this to us and we were not treated as if we were clueless either. Very nice.

Later, we took pictures of our crystals with a computer. Very sensitive and not as easy as it looked and I broke one of the capillaries so it had to be resealed. Liked the decision theatre but will write more on that in the wiki.