Abstract

In the field of protein crystallography the transfer of the protein crystals from the crystallization plate/tray, where the crystals are initially grown, to the site of analysis is a difficult and lengthy process. A new crystallization tray has been designed and 3D printed to eliminate this intermediate step which is called crystal fishing. The crystallization tray allows crystals to be grown directly on grids that are attached to the pins that are used for analysis of the crystal. The efficacy of protein crystallization using the aforementioned tray was tested in order for a protocol to be developed for potential users of the tray. Through experimentation with the proteins lysozyme and thaumatin it has been shown that the increasing the volume of crystallization solution used in the vial of the tray to maximum volume inhibits the protein crystals from drying out. Drying out can be problematic since it ruins the integrity of the crystals. Using this tray the crystals need to be able to keep for some time since the trays can be sent through the mail to be analyzed remotely. Drying out can also be prevented by equilibrating the tray with the crystallization solution for 30 minutes before loading the protein samples. Lastly, taping the tray to ensure sealing of the ports also helps to prevent the crystals from drying out.

Disciplines

Biochemistry | Structural Biology

Mentor

Silvia Russi, Aina Cohen

Lab site

SLAC National Accelerator Laboratory (SLAC)

Funding Acknowledgement

I would like to acknowledge my mentors from both SLAC and STAR, Silvia Russi, Aina Cohen, Enrique Cuellar, Larry Horvath, and Analise Elliot Heid. Thank you for all of your support and guidance this summer. I am truly grateful and am lucky to have got to work with you all. This project has been made possible with support from Chevron (www.chevron.com) and the California State University STEM Teacher Researcher Program in addition to contributions from SLAC Stanford.

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URL: http://digitalcommons.calpoly.edu/star/411

 

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