Coding
MntH

Part:BBa_K847005:Experience

Designed by: Rashmi Sharma, Vishesh Jain, Kendrick Wang   Group: iGEM12_Stanford-Brown   (2012-08-09)
Revision as of 01:01, 4 October 2012 by Kendrickwang (Talk | contribs)


Assay
Radiation
Liquid cultures of NEB5α E. coli transformed with MntH, recA, sod Cu/Zn, Dps 1 (Sod Mn and Dps 2 constructs could not be completely assembled in time), and negative control were grown up over night at 37°C. The following day, the cells were washed and resuspended in 0.9% NaCl solution. Cell concentration was then adjusted to 10^7/mL in 5mL of a glass Petri dish. Each sample was then exposed to 1.2 J/(m^2*sec) of UV-C radiation from a UV lamp for a cumulative of 0 seconds, 2 seconds, 5 seconds, 10 seconds, 20 seconds, and 30 seconds. After each exposure, a dilution spot assay was conducted to determine the final number of surviving cells. Desiccation
Liquid cultures of negative control, MntH, betAB, and otsAB transformed NEB5α E. coli were grown up over night at 37°C. After incubation, a dilution spot assay was conducted on each of the cultures to determine the density of live cells. Next, 15 5cm, round petri dishes were filled with 300uL of negative control bacteria, another 15 with transformants containing MntH, another 15 with transformants containing betAB, and another 15 with transformants containing otsAB. The petri dishes were allowed to desiccate while covered for 24 hours at 37°C while shaking. After the 24 hours period, each plate was resuspended with 1mL of fresh LB. A dilution spot assay was then conducted on each of the petri dish to determine the final density of live cells.


Results
Radiation:Characterized; however, further testing needed. Refer to: http://2012.igem.org/Team:Stanford-Brown/HellCell/Radiation. Base: Works. Refer to: http://2012.igem.org/Team:Stanford-Brown/HellCell/Desiccation

Applications of BBa_K847005

Astrobiology revolves around three central questions: "Where do we come from?", "Where are we going?", and "Are we alone?" To approach the second question, the Hell Cell subgroup of the Stanford-Brown iGEM team developed BioBricks that allow a cell to survive harsh extraterrestrial conditions. Such a toolset could create a space-ready synthetic organism to perform useful functions off-world. This gene is one of the toolset, potentially conferring radiation resistance to otherwise radiation-intolerant bacteria.

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UNIQf30eb8c5110d1a29-partinfo-00000000-QINU UNIQf30eb8c5110d1a29-partinfo-00000001-QINU