Development of a Tandem Protein Trans-Splicing System Based on Native and Engineered Split Inteins

Protein trans-splicing involving naturally or artificially split inteins results in two polypeptides being linked together by a peptide bond. While this phenomenon has found a variety of applications in chemical biology and biotechnology, precious little is known about the molecular recognition events governing the initial fragment association step. In this study, fluorescence approaches have been used to measure the dissociation constant for the Ssp DnaE split intein interaction and to determine the on and off rates of fragment association. The DnaE fragments bind with low nanomolar affinity, and our data suggest that electrostatics make an important contribution to the very rapid association of the fragments at physiological pH. This information was used to develop a tandem trans-splicing system based on native and engineered split inteins. This novel system allows the one-pot assembly of three polypeptides under native conditions and can be performed in crude cell lysates. The technology should provide a convenient approach to the segmental isotopic or fluorogenic labeling of specific domains within the context of large multidomain proteins

Reduced masses and spectroscopically determined properties of various diatomic molecules in the ground electronic state.

Reduced masses and spectroscopically determined properties of various diatomic molecules in the ground electronic state.</p

Statistical corrections of linkage data suggest predominantly regulations of gene expression-1

For each threshold of estimated using our corrected method (full pedigree analysis). The threshold for FDR = 10% is = 6.4. At this threshold we detected 22 significant linkages and expect that 2.2 are false positives. See text and Table 1 for comparison with uncorrected results.<p><b>Copyright information:</b></p><p>Taken from "Statistical corrections of linkage data suggest predominantly regulations of gene expression"</p><p>http://www.biomedcentral.com/1753-6561/1/S1/S145</p><p>BMC Proceedings 2007;1(Suppl 1):S145-S145.</p><p>Published online 18 Dec 2007</p><p>PMCID:PMC2367613.</p><p></p

Energy eigenvalues (in eV) for the various <i>n</i>_<i>r</i> and <i>l</i> quantum numbers for a few diatomic molecules by using Eq (26), where ℏ = 1.054571817×10⁻³⁴J⋅s, <i>e</i> = 1.602176634×10⁻¹⁹C, lamu = 1.66053906660×10⁻²⁷ kg (from SI Brochure 9th edition of the SI Brochure, available on the BIPM web page: www.bipm.org).

Energy eigenvalues (in eV) for the various nr and l quantum numbers for a few diatomic molecules by using Eq (26), where ℏ = 1.054571817×10−34J⋅s, e = 1.602176634×10−19C, lamu = 1.66053906660×10−27 kg (from SI Brochure 9th edition of the SI Brochure, available on the BIPM web page: www.bipm.org).</p

Comparison of the energy levels (in eV) for the various <i>n</i>_<i>r</i> and <i>l</i> quantum numbers for diatomic molecule O₂ calculated using different methods.

Comparison of the energy levels (in eV) for the various nr and l quantum numbers for diatomic molecule O2 calculated using different methods.</p

Statistical corrections of linkage data suggest predominantly regulations of gene expression-2

E findings and expected false positives using our corrected method. D, Estimated FDR curve. We identified 46 significant linkages at threshold of region-wide = 0.00036 or log() = -7.93 (FDR = 10%).<p><b>Copyright information:</b></p><p>Taken from "Statistical corrections of linkage data suggest predominantly regulations of gene expression"</p><p>http://www.biomedcentral.com/1753-6561/1/S1/S145</p><p>BMC Proceedings 2007;1(Suppl 1):S145-S145.</p><p>Published online 18 Dec 2007</p><p>PMCID:PMC2367613.</p><p></p

Shape of <i>V</i>(<i>r</i>) = <i>a</i>₋₂<i>r</i>⁻²+<i>a</i>₋₁<i>r</i>⁻¹+<i>a</i>₀ for different diatomic molecules.

Shape of V(r) = a−2r−2+a−1r−1+a0 for different diatomic molecules.</p

Minimal data set.

Finding an analytical solution to the Schrödinger equation with power function superposition potential is essential for the development of quantum theory. For example, the harmonic oscillator potential, Coulomb potential, and Klazer potential are all classed as power function superposition potentials. In this study, the general form of the power function superposition potential was used to decompose the second-order radial Schrödinger equation with this potential into the first-order Ricatti equation. Furthermore, two forms of the power function superposition potential are constructed with an exact analytical solution, and the exact bound-state energy level formula is obtained for these two potentials. Finally, the energy levels of some of the diatomic molecules were determined through calculation. And our results are actually consistent with those obtained by other methods.</div

Energy eigenvalues (in eV) for the various <i>n</i>_<i>r</i> and <i>l</i> quantum numbers for a few diatomic molecules by using Eq (36), where ℏ = 1.054571817×10⁻³⁴J⋅s, <i>e</i> = 1.602176634×10⁻¹⁹C, lamu = 1.66053906660×10⁻²⁷ kg (from SI Brochure 9th edition of the SI Brochure, available on the BIPM web page: www.bipm.org).

Energy eigenvalues (in eV) for the various nr and l quantum numbers for a few diatomic molecules by using Eq (36), where ℏ = 1.054571817×10−34J⋅s, e = 1.602176634×10−19C, lamu = 1.66053906660×10−27 kg (from SI Brochure 9th edition of the SI Brochure, available on the BIPM web page: www.bipm.org).</p

Statistical corrections of linkage data suggest predominantly regulations of gene expression-0

T panel gives the scatter plot of the scan statistics using full pedigree and sibships. For largest values of the statistics (very likely to be true positives), most of the points are above the 45° line, which suggests that full pedigree analysis provides more power than sibship analysis to detect true linkages.<p><b>Copyright information:</b></p><p>Taken from "Statistical corrections of linkage data suggest predominantly regulations of gene expression"</p><p>http://www.biomedcentral.com/1753-6561/1/S1/S145</p><p>BMC Proceedings 2007;1(Suppl 1):S145-S145.</p><p>Published online 18 Dec 2007</p><p>PMCID:PMC2367613.</p><p></p