A cryptic promoter in potato virus X vector interrupted plasmid construction

BACKGROUND: Potato virus X has been developed into an expression vector for plants. It is widely used to express foreign genes. In molecular manipulation, the foreign genes need to be sub-cloned into the vector. The constructed plasmid needs to be amplified. Usually, during amplification stage, the foreign genes are not expressed. However, if the foreign gene is expressed, the construction work could be interrupted. Two different viral genes were sub-cloned into the vector, but only one foreign gene was successfully sub-cloned. The other foreign gene, canine parvovirus type 2 (CPV-2) VP1 could not be sub-cloned into the vector and amplified without mutation (frame shift mutation). RESULTS: A cryptic promoter in the PVX vector was discovered with RT-PCR. The promoter activity was studied with Northern blots and Real-time RT-PCR. CONCLUSION: It is important to recognize the homologous promoter sequences in the vector when a virus is developed as an expression vector. During the plasmid amplification stage, an unexpected expression of the CPV-2 VP1 gene (not in the target plants, but in E. coli) can interrupt the downstream work

Normal categories of semigroup of order-preserving transformations on a finite chain

K. S. S. Nambooripad intoduced nornal categories to enable to describe the structure of regular semigroups fully. In this paper we describe the ideal categories of the regular semigroup $OX_n,$ of non-invertible order-preserving transformations on a finite chain $X_n=\{1\leq 2\leq \cdots \leq n \}$ which are normal categories. Further it is shown that the principal left ideal category of $OX_n$ as the power set category $P_o(X_n)$ of $OX_n$ and the principal right ideal category as $\prod_o(X_n)$ category of ordered partitions of $X_n$ and described the cone semigroup $T\mathscr L(OX_n)$ and prove that it is isomorphic to $OX_n.$Comment: 14 pages, 0 figure

Rancang Bangun Aplikasi Perencanaan Anggaran Biaya Tenaga Kerja pada Proyek Konstruksi Gedung

Construction project management planning will require manpower requirements, the calculation of labor cost budget, number of employees, the arrangement of activities and time required to complete each activity. So far, only managing assessments of workforce needs. This is because of the experience and do not know the technical calculations in calculating labor costs. As a result, the project failed and declining consumer confidence.Handling solutions to create budget planning application labor costs in building construction projects. The system is built to calculate the labor requirements, needs labor costs, and time required to complete the project.This application generates information report labor cost budget plan, the amount of manpower needs, and schedulling of the project. Based on the result of experiments performed, the application can help the process of planning manpower requirements, and planning labor costs in building construction projects

Local and Global Casimir Energies for a Semitransparent Cylindrical Shell

The local Casimir energy density and the global Casimir energy for a massless scalar field associated with a $\lambda\delta$-function potential in a 3+1 dimensional circular cylindrical geometry are considered. The global energy is examined for both weak and strong coupling, the latter being the well-studied Dirichlet cylinder case. For weak-coupling,through $\mathcal{O}(\lambda^2)$, the total energy is shown to vanish by both analytic and numerical arguments, based both on Green's-function and zeta-function techniques. Divergences occurring in the calculation are shown to be absorbable by renormalization of physical parameters of the model. The global energy may be obtained by integrating the local energy density only when the latter is supplemented by an energy term residing precisely on the surface of the cylinder. The latter is identified as the integrated local energy density of the cylindrical shell when the latter is physically expanded to have finite thickness. Inside and outside the delta-function shell, the local energy density diverges as the surface of the shell is approached; the divergence is weakest when the conformal stress tensor is used to define the energy density. A real global divergence first occurs in $\mathcal{O}(\lambda^3)$, as anticipated, but the proof is supplied here for the first time; this divergence is entirely associated with the surface energy, and does {\em not} reflect divergences in the local energy density as the surface is approached.Comment: 28 pages, REVTeX, no figures. Appendix added on perturbative divergence

Mathematical Model of Two Isomeric Conformations for WASP Autoinhibition

The Wiskott-Aldrich syndrome Protein (WASP) has been implicated in many diseases including Wiskott-Aldrich Syndrome (WAS) and Buruli ulcer, but no mathematical model has been developed yet to describe the kinetics/dynamics of WASP. WASP is regulated by autoinhibition. In the autoinhibited complex, intramolecular interactions with the GTPase-binding domain (GBD) occlude residues of the C terminus that regulate the Arp2/3 actin-nucleating complex. Binding of Cdc42 to the GTPase-binding domain relieves the autoinhibitory contact between the GTPase-binding domain (GBD) and the C-terminal VCA region of WASP proteins and causes a dramatic conformational change, resulting in disruption of the hydrophobic core and release of the C terminus, enabling its interaction with the actin regulatory machinery. Here we have developed a mathematical model that quantitatively describes WASP by two isomeric conformations, an active, largely unfolded conformation that is able to stimulate the Arp2/3 complex, and an inactive, folded conformation. The model invokes an intrinsic isomeric equilibrium constant  and an affinity constant  to control intramolecular contacts between the regulatory GBD and the activity-bearing VCA domain of the protein. The formulation is concentration-dependent based on steady-state equilibrium and conservation principles. By this approach we are able to quantify the fractional response of WASP against change in concentration of ligand. The model accurately predicts WASP autinhibition. The analysis confirms that WASP needs Cdc42 as an activator for maximal activation. In the absence of a ligand, WASP is regulated by the intrinsic isomeric equilibrium constant. We also find that the stability of equilibrium of the model is affected by the Cdc42 affinity of WASP.  The results further augment the understanding on the role of WASP in polymerization of actin filament and cytoskeletal rearrangement. Keywords: Wiskott - Aldrich Syndrome Protein, autoinhibition, isomeric, conformation, receptor, ligand, enzyme, protein, binding

Physical activity, exercise and low-grade systemic inflammation.

Peer Reviewe

How Does Casimir Energy Fall?

Doubt continues to linger over the reality of quantum vacuum energy. There is some question whether fluctuating fields gravitate at all, or do so anomalously. Here we show that for the simple case of parallel conducting plates, the associated Casimir energy gravitates just as required by the equivalence principle, and that therefore the inertial and gravitational masses of a system possessing Casimir energy $E_c$ are both $E_c/c^2$. This simple result disproves recent claims in the literature. We clarify some pitfalls in the calculation that can lead to spurious dependences on coordinate system.Comment: 5 pages, 1 figure, REVTeX. Minor revisions, including changes in reference