| Budget Amount *help |
¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Outline of Final Research Achievements |
The encapsulin nanocompartment from Rhodococcus erythropolis N771 was expressed and purified in wild-type and C-terminally His-tagged forms. Negative-stained transmission electron microscopy observation showed that the encapsulin monomers were assembled as a spherical particle with a diameter of 28 nm. Heterogeneous guest proteins such as enhanced green fluorescence protein and firefly luciferase were packaged into the internal cavity of the encapsulin nanocompartment by fusing the C-terminal 37-amino-acid sequence of the R. erythropolis N771 DypB peroxidase to the C-terminus. Two kinds of crystallization conditions were found for the recombinant protein. Crystals obtained using one of two conditions diffracted to 3.3Å resolution and the crystal system is trigonal, with unit-cell parameters of a = 235.5Å and α = 63.2°. A self-rotation function calculated using the collected data suggested presence of 2-, 3- and 5-fold axes in the asymmetric unit.
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