Difference between revisions of "Part:BBa K1640021"

Latest revision as of 01:45, 19 September 2015

psbWK

Sequence and Features

Overview

This biobrick contains two genes from the photosystem II complex of Chlamydomonas reinhardtii - psbW and psbK, with RBS preceeding each gene, and a terminator at the 3' end.

This part was designed in conjunction with BBa_K1640008, and together they form operon 4 in our set of photosystem II operons:

Biology & Literature

PsbW encodes psbW, a low molecular weight subunit of photosystem II (PSII), with a molecular mass of 6.1kDa in C. reinhardtii. PsbW is a transmembrane protein involved in PSII dimer-stabilisation and photoprotection. (Woolhead, Mant, Kim, Robinson, & Rodger, 2001). This protein is tightly associated with the PSII reaction centre (Shi & Schröder, 1997), with deletion studies indicating no dimeric PSII complexes present in psbW-less mutants (Shi, Lorković, Oelmüller, & Schröder, 2000). In addition, psbW-less transgenic plants have demonstrated a higher sensitivity to light-based stress (Thidholm, Shi, & Schroder, 2001).

PsbK encodes psbK, a low molecular weight subunit of PSII involved in stabilising the complex, however the necessity of psbK varies between species. In psbK-less C. reinhardtii, the PSII complex was destabilised, with >10% of PSII complex observed, compared to the wild type (Takahashi, Matsumoto, Goldschmidt-Clermont, & Rochaix, 1994). PsbK-less Synechocytis sp., however, only experienced a marginally reduced rate of electron transport and growth (Ikeuchi et al., 1991). In C. reinhardtii, psbK is bound to CP43 (Sugimoto & Takahashi, 2001).

Protein information

psbW

mass: 9.2kDa

sequence: MATTVRSEVAKKVAMLSTLPATLAAHPAFALVDERMNGDGTGRPFGVNDPVLGWVLLGVFGTMWAIWFIGQKDLGDFEDADDGLKL

psbK

mass: 5.0kDa

sequence: MTTLALVLAKLPEAYAPFAPIVDVLPVIPVFFILLAFVWQAAVSFR

Part verification

Visualisation of psbWK showing expected banding is shown in the following gel image, bottom row first set of lanes, with left lane part showing EcoRI digest, right lane showing EcoRI + PstI double digest. This part has been sequenced to confirm design with final biobrick.

References

Ikeuchi, M., Eggers, B., Shen, G., Webber, A., Yu, J., Hirano, A., . . . Vermaas, W. (1991). Cloning of the psbK gene from Synechocystis sp. PCC 6803 and characterization of photosystem II in mutants lacking PSII-K. Journal of Biological Chemistry, 266(17), 11111-11115.

Shi, L.-X., Lorković, Z. J., Oelmüller, R., & Schröder, W. P. (2000). The low molecular mass PsbW protein is involved in the stabilization of the dimeric photosystem II complex in Arabidopsis thaliana. Journal of Biological Chemistry, 275(48), 37945-37950.

Shi, L.-X., & Schröder, W. (1997). Compositional and topological studies of the PsbW protein in spinach thylakoid membrane. Photosynthesis Research, 53(1), 45-53. doi:10.1023/A:1005830405809

Sugimoto, I., & Takahashi, Y. (2001). Localization of a small chloroplast-encoded polypeptide PsbK in photosystem II core complex. Science Access, 3(1).

Takahashi, Y., Matsumoto, H., Goldschmidt-Clermont, M., & Rochaix, J.-D. (1994). Directed disruption of the Chlamydomonas chloroplast psbK gene destabilizes the photosystem II reaction center complex. Plant molecular biology, 24(5), 779-788.

Thidholm, E., Shi, L.-X., & Schroder, W. (2001). The PsbW-protien; Its location and involvement in photoinhibition. Science Access, 3(1).

Woolhead, C. A., Mant, A., Kim, S. J., Robinson, C., & Rodger, A. (2001). Conformation of a purified “spontaneously” inserting thylakoid membrane protein precursor in aqueous solvent and detergent micelles. Journal of Biological Chemistry, 276(18), 14607-14613.