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A highly efficient system was developed for the expression of foreign genes in Chlorella ellipsoidea cells. The effect of five promoters on the expression efficiency of beta-glucuronidase (GUS) gene was evaluated by transient expression of the UidA gene. Among these promoters, Ubiquitin-omega was found to be the most efficient and was selected to drive the expression of foreign genes in Chlorella cells. A gene encoding the mature rabbit neutrophil peptide-1 (NP-1) was introduced into the cells. Integration of the gene for NP-1 into the Chlorella genome was confirmed by PCR and Southern blot analysis. In, vitro anti-microbial testes demonstrated the expression of biologically active NP-1 by the transgenic Chlorella cells.

In an exploratory study, using the tadpole improved clover Wilson quark action on small, coarse and anisotropic lattices, the ππ scattering length in the I = 2 channel is calculated within quenched approximation. A new method is proposed which enables us to make chiral extrapolation of our lattice results without calculating the decay constant on the lattice. Finite volume and finite lattice spacing errors are analyzed and the results are extrapolated towards the infinite volume and continuum limit. Comparisons of our lattice results with the new experiment and the results from Chiral Perturbation Theory are made. Good agreements are found.

The lowest-lying glueballs are investigated in lattice QCD using N f = 2 clover Wilson fermion on anisotropic lattices. We simulate at two different and relatively heavy quark masses, corresponding to physical pion mass of mπ ∼ 938 MeV and 650 MeV. The quark mass dependence of the glueball masses have not been investigated in the present study. Only the gluonic operators built from Wilson loops are utilized in calculating the corresponding correlation functions. In the tensor channel, we obtain the ground state mass to be 2.363(39) GeV and 2.384(67) GeV at mπ ∼ 938 MeV and 650 MeV, respectively. In the pseudoscalar channel, when using the gluonic operator whose continuum limit has the form of ijk T rBiDjB k , we obtain the ground state mass to be 2.573(55) GeV and 2.585(65) GeV at the two pion masses. These results are compatible with the corresponding results in the quenched approximation. In contrast, if we use the topological charge density as field operators for the pseudoscalar, the masses of the lowest state are much lighter (around 1GeV) and compatible with the expected masses of the flavor singlet qq meson. This indicates that the operator ijk T rBiDjB k and the topological charge density couple rather differently to the glueball states and qq mesons. The observation of the light flavor singlet pseudoscalar meson can be viewed as the manifestation of effects of dynamical quarks. In the scalar channel, the ground state masses extracted from the correlation functions of gluonic operators are determined to be around 1.4-1.5 GeV, which is close to the ground state masses from the correlation functions of the quark bilinear operators. In all cases, the mixing between glueballs and conventional mesons remains to be further clarified in the future.

Roundup-Ready soybeans have been genetically modified to resist the effects of the herbicidal glyphosate and have become the most prevalent transgenic crop in the world. In this work, Roundup-Ready soybeans were used as raw material to study the effects of critical processing procedures such as grinding, cooking, blending, homogenization, sterilization, and spray-drying on the length of DNA fragments of an endogenous gene (lectin) and an exogenous gene (epsps) examined in material from three soybean foods of bean curd, soy milk, and soy powder and from samples taken during their processing. The results showed that various processing procedures caused degradations of both the endogenous and exogenous genes to different degrees. In the grinding procedure, endogenous gene DNA was degraded from 1883 to approximately 836 bp, and exogenous gene DNA was degraded from 1512 to approximately 408 bp. In the blending and squeeze-molding procedures, exogenous gene DNA was also degraded from about 408 to 190 bp, but there was no obvious action on the endogenous gene. After the endogenous and exogenous genes had been degraded to some degree, such as 836 and 408 bp, respectively, they were not evidently affected by cooking procedure at 100 degrees C for 15 min. However, the endogenous gene was further considerably degraded from around 836 to 162 bp in the sterilization procedure at 121 degrees C for 30 s. The effect of the homogenization step on endogenous and exogenous genes was similar to that of the cooking procedure. The coagulation procedure, principally a biochemical reaction, did not greatly affect the exogenous gene but did affect endogenous gene, reducing DNA size from about 836 to 407 bp. Furthermore, the spray-drying procedure, a process of physical shearing, high temperature, and sudden high pressure, distinctly caused degradation of both the lectin and epsps genes, rapidly decreasing the sizes from about 836 to 162 bp for the endogenous gene and from about 408 to 190 bp for the exogenous gene.

Synechocystis sp. PCC 6803 (hereafter Synechocystis) is a model cyanobacterium and has been used extensively for studies concerned with photosynthesis and environmental adaptation. Although dozens of protein kinases and phosphatases with specificity for Ser/Thr/Tyr residues have been predicted, only a few substrate proteins are known in Synechocystis. In this study, we report 194 in vivo phosphorylation sites from 149 proteins in Synechocystis, which were identified using a combination of peptide pre-fractionation, TiO(2) enrichment and liquid chromatograpy-tandem mass spectrometry (LC-MS/MS) analysis. These phosphorylated proteins are implicated in diverse biological processes, such as photosynthesis. Among all identified phosphoproteins involved in photosynthesis, the β subunits of phycocyanins (CpcBs) were found to be phosphorylated on Ser22, Ser49, Thr94 and Ser154. Four non-phosphorylated mutants were constructed by using site-directed mutagenesis. The in vivo characterization of the cpcB mutants showed a slower growth under high light irradiance and displayed fluorescence quenching to a lower level and less efficient energy transfer inside the phycobilisome (PBS). Notably, the non-phosphorylated mutants exhibited a slower state transition than the wild type. The current results demonstrated that the phosphorylation status of CpcBs affects the energy transfer and state transition of photosynthesis in Synechocystis. This study provides novel insights into the molecular mechanisms of protein phosphorylation in the regulation of photosynthesis in cyanobacteria and may facilitate the elucidation of the entire regulatory network by linking kinases to their physiological substrates.

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In this exploratory lattice study, low-energy scattering of the (DD * ) ± meson system is analyzed using lattice QCD with N f = 2 twisted mass fermion configurations with three pion mass values. The calculation is performed within single-channel Lüscher's finite-size formalism. The threshold scattering parameters, namely the scattering length a0 and the effective range r0, for the s-wave scattering in J P = 1 + channel are extracted. For the cases in our study, the interaction between the two charmed mesons is weakly repulsive. Our lattice results therefore do not support the possibility of a shallow bound state for the two mesons for the pion mass values we studied. This calculation provides some useful information on the nature of the newly discovered resonance-like structure Zc(3900) by various experimental groups.