Isolation, properties and spatial site analysis of gamma subunits of B-phycoerythrin and R-phycoerythrin

Polysiphonia urceolata R-phycoerythrin and Porphyridium cruentum B-phycoerythrin were degraded with proteinaseK, and then the nearly native gamma subunits were isolated from the reaction mixture. The process of degradation of phycoerythrin with proteinaseK showed that the gamma subunit is located in the central cavity of (alpha beta)(6) hexamer of phycoerythrin. Comparative analysis of the spectra of the native phycoerythrin, the phycoerythrin at pH 12 and the isolated gamma subunit showed that the absorption peaks of phycoerythrobilins on alpha or beta subunit are at 535 nm (or 545 nm) and 565 nm, the fluorescence emission maximum at 580 nm; the absorption peak of phycoerythrobilins on the isolated gamma subunit is at 589 nm, the fluorescence emission peak at 620 nm which overlaps the absorption maximum of C-phycocyanin and perhaps contributes to the energy transfer with high efficiency between phycoerythrin and phycocyanin in phycobilisome; the absorption maximum of phycourobilin on the isolated gamma subunit is at 498 nm, which is the same as that in native phycoerythrin, and the fluorescence emission maximum at 575 nm.

Spectroscopic properties of the C-phycocyanin-allophycocyanin conjugate and the isolated phycobilisomes from Spirulina platensis

C-phycocyanin (CPC) and allophycocyanin (APC) were purified from Spirulina platensis, then the CPC was attached covalently to the APC by reacting their epsilon-amino groups. The excitation energy could be transferred from the CPC to the APC in the CPC-APC conjugate. Intact phycobilisomes (PBS), consisting of CPC, APC, colourless linker polypeptides, and APC B or L-cm, were isolated from S. platensis. Spectroscopic properties of the isolated PBSs kept at 20 degrees C for various times showed that the connection between the APC and the APC B or L-cm was looser than that between the CPC and the APC in the isolated PBSs. The CPC-APC conjugate was more stable than the isolated PBSs, and the linker polypeptides had a minor influence on the excitation energy transfer characteristic between different phycobiliproteins in the PBS.

Biosynthesis of fluorescent cyanobacterial allophycocyanin trimer in Escherichia coli

Allophycocyanin (APC), a cyanobacterial photosynthetic phycobiliprotein, functions in energy transfer as a light-harvesting protein. One of the prominent spectroscopic characteristics of APC is a strong red-shift in the absorption and emission maxima when monomers are assembled into a trimer. Previously, holo-APC alpha and beta subunits (holo-ApcA and ApcB) were successfully synthesized in Escherichia coli. In this study, both holo-subunits from Synechocystis sp. PCC 6803 were co-expressed in E. coli, and found to self-assemble into trimers. The recombinant APC trimer was purified by metal affinity and size-exclusion chromatography, and had a native structure identical to native APC, as determined by characteristic spectroscopic measurements, fluorescence quantum yield, tryptic digestion analysis, and molecular weight measurements. Combined with results from a study in which only the monomer was formed, our results indicate that bilin synthesis and the subsequent attachment to apo-subunits are important for the successful assembly of APC trimers. This is the first study to report on the assembly of recombinant ApcA and ApcB into a trimer with native structure. Our study provides a promising method for producing better fluorescent tags, as well as a method to facilitate the genetic analysis of APC trimer assembly and biological function.

Evolutionary analysis of phycobiliproteins: Implications for their structural and functional relationships

Phycobiliproteins, together with linker polypeptides and various chromophores, are basic building blocks of phycobilisomes, a supramolecular complex with a light-harvesting function in cyanobacteria and red algae. Previous studies suggest that the different types of phycobiliproteins and the linker polypeptides originated from the same ancestor. Here we retrieve the phycobilisome-related genes from the well-annotated and even unfinished cyanobacteria genomes and find that many sites with elevated d(N)/d(S) ratios in different phycobiliprotein lineages are located in the chromophore-binding domain and the helical hairpin domains (X and Y). Covariation analyses also reveal that these sites are significantly correlated, showing strong evidence of the functional-structural importance of interactions among these residues. The potential selective pressure driving the diversification of phycobiliproteins may be related to the phycobiliprotein-chromophore microenvironment formation and the subunits interaction. Sites and genes identified here would provide targets for further research on the structural-functional role of these residues and energy transfer through the chromophores.

电离层行进式扰动的GPS台网监测研究

In this dissertation, we investigated two types of traveling ionospheric disturbances (TIDs)/gravity waves (GWs) triggered separately by auroral energy input during super geomagnetic storms and solar terminator (ST) under quiet geomagnetic conditions (kp<3+) using TEC measurements from the global network of GPS receivers. Research into the generation and propagation of TIDs/GWs during storms greatly enhance our understandings on the evolution processes of energy transportation from the high-latitude’s magnetosphere to the low-latitude ionosphere and the conjugated effect of TIDs propagation between the northern and southern hemispheres. Our results revealed that the conjugacy of propagation direction between the northern and southern hemispheres was subject to the influence of Coriolis force. We also figure out the evolution processes of ionospheric disturbances at the global scale. These are important topics that had not been well addressed previously. In addition, we also obtained thee wave structures of medium scale TIDs excited by the solar terminator (ST) moving over the northern America and physical mechanisms involved. Our observations confirm that the ST is a stable and repetitive source of ionospheric wave disturbances and the evidence of solar terminator generated disturbances has been demonstrated experimentally via the GPS TEC measurement. The main researches and results of this dissertation are as follows. First, the global traveling ionospheric disturbances (TIDs) during the drastic magnetic storms of October 29–31, 2003 were analyzed using the Global Position System (GPS) total electron content (TEC) data observed in the Asian-Australian, European and North American sectors. We collected the most comprehensive set of the TEC data from more than 900 GPS stations on the International GNSS Services (IGS) website and introduce here a strategy that combines polynomial fitting and multi-channel maximum entropy spectral analysis to obtain TID parameters. Moreover, in collaboration with my thesis advisor, I have developed an imaging technique of 2-dimensional map of TIDs structures to obtain spatial and temporal maps of large scale traveling ionospheric disturbances (LSTIDs). The clear structures of TEC perturbations map during the passage of TIDs were displayed. The results of our study are summarized as follows: (1) Large-scale TIDs (LSTIDs) and medium-scale TIDs (MSTIDs) were detected in all three sectors after the sudden commencement (SC) of the magnetic storm, and their features showed longitudinal and latitudinal dependences. The duration of TIDs was longer at higher latitudes than at middle latitudes, with a maximum of about 16 h. The TEC variation amplitude of LSTIDs was larger in the North American sector than in the two other sectors. At the lower latitudes, the ionospheric perturbations were more complicated, and their duration and amplitude were relatively longer and larger. (2) The periods and phase speeds of TIDs were different in these three sectors. In Europe, the TIDs propagated southward; in North America and Asia, the TIDs propagated southwestward; in the near-equator region, the disturbances propagated with the azimuth (the angle of the propagation direction of the LSTIDs measured clockwise from due north with 0°) of 210° showing the influence of Coriolis force; in the Southern Hemisphere, the LSTIDs propagated conjugatedly northwestward. Both the southwestward and northeastward propagating LSTIDs are found in the equatorial region. These results mean that the Coriolis effect cannot be ignored for the wave propagation of LSTIDs and that the propagation direction is correlated with the polar magnetic activity. (3) The day (day of year: 301) before the SC (sudden commencement) of magnetic storm, we observed a sudden TEC skip disturbances (±10 TECU). It should be a response for the high flux of proton during the solar flare event, but not the magnetic storms. Next, the most comprehensive and dense GPS network’s data from North-America region were used in this paper to analyze the medium scale traveling ionospheric disturbances (MSTIDs) which were generated by the moving solar terminator during the quiet days in 2005. We applied the multi-channel maximum entropy spectral analysis to calculated TID parameters, and found that the occurrence of ST-MSTIDs depends on the seasonal variations. The results of our study are summarized as follows: (1) MSTIDs stimulated by the moving ST (ST-MSTIDs) are detected at mid-latitudes after the passage of the solar terminator with the life time of 2～3 hours and the variation amplitude of 0.2～0.8 TECU. Spectral analysis indicated that the horizontal wavelength, average period, horizontal phase velocity of the MSTIDs are around 300±150 km，150±80 m/s and 25±15 min, respectively. In addition, ST-MSTIDs have wave fronts elongating the moving ST direction and almost parallel to ST. (2) The statistical results demonstrate that the dusk MSTIDs stimulated by ST is more obvious than the dawn MSTIDs in summer. On the contrary, the more-pronounced dawn MSTIDs occurs in winter. (3) Further analysis indicates that the seasonal variations of ST-MSTIDs occurrence frequency are most probably related to the seasonal differences of the variations of EUV flux in the ionosphere region and recombination process during sunrise and sunset period at mid-latitudes. Statistical study of occurrence characteristics of TIDs using the GPS network in North-American and European during solar maximum, In conclusion, statistical studies of the propagation characteristics of TIDs, which excited by the two common origins including geomagnetic storms and moving solar terminator, were involved with global GPS TEC databasein this thesis. We employed the multichannel maximum entropy spectral analysis method to diagnose the characteristics of propagation and evolvement of ionospheric disturbances, also, the characteristics of their regional distribution and climatological variations were revealed by the statistic analysis. The results of these studies can improve our knowledge about the energy transfer in the solar-terrestrial system and the coupling process between upper and lower atmosphere (thermosphere-ionosphere-mesosphere). On the other hand, our results of the investigation on TIDs generated by particular linear origin such as ST are important for developing ionospheric irregularity physics and modeling the transionosphere radio wave propagation. Besides, the GPS TEC representation of the ST-generated ionospheric structure suggests a better possibility for investigating this phenomenon. Subsequently, there are scientific meaning of the result of this dissertation to deeply discuss the energy transfer and coupling in the ionosphere, as well as realistic value to space weather forecast in the ionosphere region.

A homogeneous DNA hybridization system by using a new luminescence terbium chelate

Homogeneous DNA hybridization assay based on the luminescence resonance energy transfer (LRET) from a new luminescence terbium chelate, N,N,N-1,N-1-[2,6-bis(3'-aminomethyl-1'-pyrazolyl)-4-phenylpyridine]tetrakis(acetic acid) (BPTA)-Tb3+ (lambda(ex) = 325 nm and lambda(em) = 545 nm) to an organic dye, Cy3 (A,. = 548 nm and A,. = 565 nm), has been developed. In the system, two DNA probes whose sequences are complementary to the two different consecutive sequences of a target DNA are used; one of the probes is labeled with the Tb3+ chelate at the T-end, and the other is with Cy3 at the 5'-end. Labeling of the Tb3+ chelate is accomplished via the linkage of a biotin-labeled DNA probe with the Tb3+ chelate-labeled streptavidin. Strong sensitized emission of Cy3 was observed upon excitation of the Tb3+ chelate at 325 run, when the two probe DNAs were hybridized with the target DNA. The sensitivity of the assay was very high compared with those of the previous homogeneous-format assays using the conventional organic dyes; the detection limit of the present assay is about 30 pM of the target DNA strand.

Application of dynamical Lie algebraic method to atom-diatomic molecule scattering

The dynamical Lie algebraic approach developed by Alhassid and Levine combined with intermediate picture is applied to the study of translational-vibrational energy transfer in the collinear collision between an atom and an anharmonic oscillator. We find that the presence of the anharmonic terms indeed has an effect on the vibrational probabilities of the oscillator. The computed probabilities are in good agreement with those obtained using exact quantum method. It is shown that the approach of dynamical Lie algebra combining with intermediate picture is reasonable in the treating of atom-anharmonic oscillator scattering.