Structural analysis of the promoter of tomato 1-aminocyclopropane-1-carboxylate synthase 6 gene(Le-ACS6)

Ethylene plays an important role in the regulation of many growth and developmental processes of higher plants. In tomato,Le-ACS6,a member of the ACC synthase multigene family involved in system 1 ethylene biosynthesis during fruit ripening,is subject to negative feedback regulation by ethylene. To identify the cis-elements that are responsible for the negative feedback control,we established an in vitro transient assay system employing particle bombardment on mature-green tomato fruit pericarp to examine the expression of a luciferase(LUC) reporter gene driven by a 5′-serially deleted Le-ACS6 promoter. The results localized putative cis-elements required for negative ethylene-response between -347 and -266 upstream from the translational start site ATG. Several lines of stable transformation of the Le-ACS6 promoter and GUS reporter fusion gene containing internal deletion from -347 to -266 were generated. The expression pattern of the GUS reporter showed that removal of the nucleotides from -347 to -266 completely eliminated the response of the Le-ACS6 promoter to exogenous ethylene.     

Analysis of the Cotton E6 Promoter

An E6 gene from sea island cotton （Gossypium barbadense） was expressed specifically in cotton fiber cells to transfer functions to cultivated species for better transgenic engineering. The regulatory activity of the E6 promoter region was then studied by isolating a 614-bp fragment of the 5＇-flanking region from upland cotton （Gossypium hirsutum CR1-12） to produce a green fluorescent protein （GFP） reporter construct for analysis of tissue-specific expression in transgenic tobacco seedlings. Fluorescent analyses indicate that the relatively short E6 promoter is sufficient to direct green fluorescent protein expression specifically in the leaf trichomes （hair cells） of the transgenic tobacco plants. As cotton fibers are also unicellular trichomes that differentiate from epidermal cells of developing cotton ovules, the result suggests that the relatively short E6 promoter can serve as a fiber-specific expression promoter for genetic engineering to improve cotton fiber quality.     

Evaluation and characterization of an endosperm-specific sbeⅡa promoter in wheat

Starch,the main component of the wheat grain,is the product of a complex biochemical pathway. The sbeⅡα gene plays a key role in controlling the synthesis of starch, in particular, the biosynthesis of amylopectin,in maturing wheat grain.To investigate its regulatory mechanisms and endosperm-specific expression pattern, the sbeⅡα promoter (3094 bp in length) was cloned using APCR and sequenced.The effect of a series of deletions was studied using a GUS transient assay system. Results showed that the 3094 bp sequence (sbe.g construct) exhibited full stable promoting activity and that the activities of 5′ or 3′ deletions reduced levels of GUS expression. Some constructs with internal deletions showed only weak activity, however,sbe.e, with a deletion from -1579—--1210 bp resulted in higher levels of expression than the full-length promoter sequence, sbe.g. This indicates that motifs such as the -300 bp element, G-box and/or P-box act as positive elements and are necessary in determining the promoter‘s endosperm-specific pattern and that negative repressor elements or motifs may also be present within the -1579—-1210 bp sequence. The age of wheate ndosperm tissue used in the GUS-transient assay system is shown to be of significant importance.     

Promoter of soybean early nodulin gene<Emphasis Type="Italic">enod2B</Emphasis> is induced by rhizobial Nod factors in transgenic rice

Nod factors, which are signaling molecules produced by Rhizobia, are the principal determinants of host specificity in Rhizobium-legume symbiosis. Nod factors can elicit a number of characteristic developmental responses in the roots of legumes, such as depolarization of the membrane potential in epidermal cells, specific expression of early nodulin genes and changes in the flux of calcium in root hairs, deformation of root hairs, cell division in the root cortex and formation of the nodule primordinm. Whether the rice plant can respond to signaling molecules (i.e. Nod factors) is an important question, as it could establish the potential for symbiotic nitrogen fixation in rice. The promoter of the soybean (Glycine max) early nodulin gene Gmenod2B fused to the β-glucuronidase (GUS) reporter gene was used as a molecular marker to explore whether Nod factors can be recognized by rice cells as signaling molecules. Transgenic rice plants harboring the chimeric gene Gmenod2BP-GUS were obtained via an Agrobacterium tumefaciens-mediated system. NodNGR factors produced by a broad-host-range Rhizobium strain NGR234(pA28) were used as probes to investigate the activity of the Gmenod2B promoter in rice. Our results showed that the early nodulin gene Gmenod2B promoter was induced by NodNGR factors in transgenic rice, and that it was specifically expressed in rice plant roots. Moreover, GUS gene expression driven by the Gmenod2B promoter in transgenic rice was regulated by nitrogen status. These findings indicated that rice possessed the ability to respond to Nod factor signals, and that this signal transduction system resulted in activation of the Gmenod2B promoter. Thus, we predict that the Nod-factor inducible nodulin expression system, which is similar to Rhizobium-legume symbiosis, may also exist in rice.     

Isolation of a pollen specific promoter in tritordeum

The promoter is a cis-acting element in regulating gene expression. A promoterless plasmid containing UidA gene was transformed into tritordeum by barmbadment. Histochemical analysis of various tissues in transgenic tritordeum was carried to examine tissue-specific expression of GUS(beta-glucuronidase) activity. The pollen-specific promoter was trapped and identified successfully in a transformant line. PCR(polymerase chain reaction) method was used to isolate this pollen-specific promoter. By sequencing and analyzing the amplified fragment from PCR, a part of UidA gene and a flanking sequence were obtained. Some essential elements of plant promoters were found in the sequence. To determine the function of it, the cloned fragment was fused with UidA gene, then cloned and transformed into Triticum durum. The transgenic plant transformed by this vector showed GUS expression only in pollen. Therefore a pollen-specific promoter was isolated successfully.     

Functional analysis of the larval serum protein gene promoter from silkworm <Emphasis Type="Italic">Bombyx mori.</Emphasis>

The regulation region of larval serum protein gene, Bombyx mori. (BmLSP), consisting of the first intron, the first exon, the central promoter region and 5′-upstream region, is cloned from genomic DNA from the silkworm va-riety of Suju譓inghu. Using PCR and restriction endonu-clease methods, a series of luciferase reporter plasmids, driven by different length of BmLSP promoters, are con-structed. Via the transient expression system in BmN cells, the effects of the regulation elements and foreign insect hor-mones on the BmLSP promoter activity are investigated. The results demonstrate that the promoter activity of BmLSP is 5.8- or 4.4-fold higher than that of BmLSPs whose first in-tron or the element in 5′-upstream region harboring the homologous sequence with the first intron of light-chain fib-roin gene (EHIF) is deleted, respectively, suggesting that both the first intron and EHIF contain the main positive cis-acting elements. However, the inactive mariner transposable ele-ment (MTE) in 5′-upstream region presents a negative effect. Furthermore, the effects of juvenile hormone analogue (JHA) on the BmLSP promoter activity show a typical dose-dependent manner, that is, low concentration treat-ments increase the BmLSP promoter activity and high con-centration treatments decrease it. Meanwhile, insect ecdy-sone (MH) treatments present no significant effect.     

A novel element (NIRS) participates in the regulation ofinterleukin 2 receptorαgene expression

A novel element at -153/- 143 bp in the interleukin 2 receptor α(IL-2Rα) gene has been coined as NRE-inverse repeat sequence (NIRS) due to its inversely repeated to the known negative regulatory element (NRE) further upstream of the gene. In order to explore the role of NIRS in the expression of IL-2Rαgene,luciferase reporter plasmids driven by 4 individually deleted IL-2Rα genes promoter regions were constructed. Transfection of the reporter plasmids into Jurkat cells and HeLa cells respectively, we found that both NIRS and NRE were critical for repressing the constitutive expression of IL-2Rα gene and were also necessary for promoter activity induced by PHA. EMSA results showed that double-stranded NRE- and NIRS-binding proteins existed in both HeLa cells and Jurkat cells. However, single-stranded NIRS- and NRE-binding protein was only found in HeLa cells. Interestingly, the supershift band showed up in EMSA system with Jurkat cells (no matter whether activated or not) adding to the cell lysate of HeLa cells. UV-crosslinking showed a double stranded NRE- and NIRS-binding protein p83 in both Jurkat cells and HeLa cells. Our results suggest that trans-acting factors play a key role in regulating promoter activity of IL-2Rα gene by interacting with double or single stranded NRE and/or NIRS selectively in different cells.     

Compression behavior and equation of state of Ni77P23 amorphous alloy

The compression behavior of Ni77P23 amorphous alloy is investigated at room temperature in a diamond-anvil cell instrument using insitu high pressure energy dispersive X-ray diffraction with a syn- chrotron radiation source. The equation of state is determined by fitting the experimental data accord- ing to Birch-Murnaghan equation: -ΔV/V0=0.08606P-3.2×10-4P2 5.7×10-6P3. It is found that the structure of Ni77P23 amorphous alloy is stable under pressures up to 30.5 GPa.     

Pollen flux and vertical dispersion in coniferous and deciduous broadleaved mixed forest in the Changbai Mountains

The pollen flux in coniferous and deciduous broadleaved mixed forest in the Changbai Mountains is presented in one year's experiments. The results indicate that arboreal pollen percentages are more than 65% and pollen flux is higher than 5000 grain·cm-2·a-1,while less than 2% and lower than 1000 grain·cm-2·a-1 for shrubby pollen,and less than 20% and lower than 3000 grain·cm-1·a-1 for herbaceous pollen for most samples at different heights. The pollen assemblages are similar to those in the samples under 8 m height where Pinus and Ouercus are dominant and followed by the few non-local pollen types,and Fraxinus percentages are high with a few non-local pollen types at 16 m to 32 m height as well as non-local pollen clearly increased at 40 m height. Comparisons between pollen assemblages and vegetation composition suggest that similarities are higher for pollen trap samples than for surface moss samples.     

Investigation on chain structure of LLDPE obtained by ethylene in-situ copolymerization with DSC and XRD

Thermal segregations of LLDPE were treated with successive self-nucleation/annealing (SSA) on dif-ferential scanning calorimetry (DSC). Information on molecular heterogeneity of LLDPE was obtained. After SSA was treated, the multiple endothermic peaks were observed in the DSC thermograms during heating experiment. It is obtained that the thickness of different lamellas formed by segments of vari-ous lengths was 4―10 nm. X-ray diffraction (XRD) results showed that the crystallites dimensions of various reflections were about several dozens of nanometers. The ethylene/α-olefin copolymers and the copolymer via in-situ copolymerization were similar to each other for molecular heterogeneity and XRD characteristics, which revealed that it was possible to use the ethylene/α-olefin copolymers to simulate the copolymer via in-situ copolymerization of ethylene to simplify the complexity of the structure of the ethylene in-situ copolymer.     

SCFP,a novel fiber-specific promoter in cotton

To investigate the expression pattern of GhSCFP which was isolated from cotton fiber cDNA library, a 1006 bp upstream fragment of the gene was cloned by chromosome walking and fused to GUSand GFP respectively. Histochemical GUS and GFP fluorescence analysis revealed that the expression of the report genes driven by the promoter sequence was detectable only in outer layer cells during the seed development in the transgentic tobaccos. In transgenic cotton, strong GUS activity was observed in spherical protrusions on 0 dpa （days post anthesis） ovule surface, and in the 2-36 dpa fiber cells, while no GUS signals were detected in the root, leaves, stem, corolla, anther and stigma. Our data demonstrated that GhSCFP upstream sequence is a cotton fiber-specific promoter and this promoter will be useful in the molecular research on fiber cell development and in cotton fiber improvements by genetic modification.     

Southwest monsoon changes indicated by oxygen isotope of ostracode shells from sediments in Qinghai Lake since the late Glacial

The δ 18O records of ostracode shells in sediments of core QH-2000 from Qinghai Lake can be used as a better proxy to reflect monsoon changes. Low monsoon precipitation between 17.5 and 11 cal. ka BP is indicated by positive δ 18O values averaging 2.37‰. A fast shift in δ 18O from positive at 11 cal. ka BP to negative at 10 cal. ka BP indicates sharp increase of monsoon precipitation. An interval of generally high monsoon precipitation is observed between 10 and 6 cal. ka BP with δ 18O values averaging -2.15‰. Decrease of monsoon precipitation between 6 and 2.5 cal. ka BP is indicated by positive δ 18O values. δ 18O keeps positive values averaging 3.0‰ between 2.5 and 0 cal. ka BP suggesting low high monsoon precipitation. The climatic changes indicated by δ 18O records of ostracode shells in sedi- ments of core QH-2000 from Qinghai Lake and our broader regional comparison show that the climate in Qinghai Lake since the late Glacial is probably controlled by southwest monsoon other than southeast monsoon.     

Temperature dependence of densities of Sb and Bi melts

The densities of Sb and Bi melts were investigated by an improved Archimedean method. The results show that the density of the Sb melt decreases linearly with increasing temperature,but the density of the Bi melt firstly increases and then decreases as the temperature increases. There is a maximum density value of 10.002 g/cm3 at 310 ℃ ,about 39℃ above the melting point. The temperature depend-ence of the Sb melt is well fitted with the expression ρ =6.8590-5.8105×10-4T,and that of the Bi melt is fitted with ρ =10.3312-1.18×10-3T. The results were discussed from a microstructure viewpoint.     

Dead lithium phase investigation of Sn-Zn alloy as anode materials for lithium ion battery

In this work, based on First-principle plane wave pseudo-potential method, we have carried out an in-depth study on the possible dead lithium phase of Sn-Zn alloy as anode materials for lithium ion batteries. Through investigation, we found that the phases Li_xSn₄Zn₄(x = 2, 4, 6, 8) contributed to reversible capacity, while the phases Li_xSn₄Zn_8−(x−4)(x = 4.74, 7.72) led to capacity loss due to high formation energy, namely, they were the dead lithium phases during the charge/discharge process. And we come up with a new idea that stable lithium alloy phase with high lithiation formation energy (dead lithium phase) can also result in high loss of active lithium ion, besides the traditional expression that the formation of solid electrolyte interface film leads to high capacity loss. Supported by the National Natural Science Foundation of China (Grant No. 50771046), Natural Science Foundation of Guangdong Province (Grant No. 05200534), Key Projects of Guangdong Province and Guangzhou City (Grant Nos. 2006A10704003 and 2006Z3-D2031) and China Postdoctoral Science Foundation (Project No. 20080440764)     

Composition dependence of viscosity for Al（1-x）Mgx（0≤x≤0. 10） alloys

Viscosities of molten Al（1-x）Mgx（0≤x≤0. 10 alloys have been measured in the temperature range of 973 K--1173 K by a torsional oscillation cup method. The viscosity dependence on temperature for Al（1-x）Mgx（0≤x≤0. 10 melts obeys Arrhenius equation. The viscosity increases with increasing magnesium concentration in the investigated system. There is an important relationship between viscosity and its glass-forming ability for metallic melt.     

Synthesis and X-ray crystal structure of a new manganese 18-metallacrown-6

0　IntroductionThemetalionsinmetallamacrocylesformedbysupramolecularself assemblycantaketrigonal,square planar,andtetrahe dralconfiguration[1 ,2 ] .Metallacrownisaspecialclassofmetalla macrocyle,theyareanalogoustocrownethersinbothstructureandfunctionexceptthatthemetalionsarenowtakingthepositionsofcoordinationatomsincrownether[3,4] .Themetallacrownswithdifferentnumberof [M—N—O]repeatunithavedifferentcavitysizes,suchas 9 MC 3[5 7] ,12 MC 4 [4,8 1 4 ] and 15 MC 5 [1 5] .Thereportedazameta…