转基因烟草中的海藻糖测定

The \%E.coli\% trehase synthalose gene(\%otsA\%) was transferred into \%Nicotiana tabacum\% mediated by \%Agrobacterium\%, but the method for detecting low concentration of trehalose in transgenic plant was not available.The high performance liquid chromatograph(HPLC) with evaporative light\|scatting detector (ELSD) using water:methyl cyanide(1∶2\^6 v/v) as mobile phase was established in this work. An ODS column Zorbax RX\|SIL was employed. The trehalose detection limits of ELSD was 5mg/L. From the linear…     

中国家蚕抗菌肽人工合成类CMⅣ基因在甜菜夜蛾虫体中的表达

将人工合成的中国家蚕抗菌肽类CMⅣ基因与抗菌肽信号肽基因连接 ,经EcoRⅠ、HindⅢ双酶切后 ,克隆于pFASTBacⅠ的EcoRⅠ、HindⅢ酶切位点之间 ,得到重组转座载体pFASTBac ABP ,经测序证明阳性克隆正确。将重组转座载体转化HD1 0Bac大肠杆菌 ,得到重组Bacmid ABP。将重组Bacmid转染sf2 1细胞及感染甜菜夜蛾 (Laphygmaexigua)幼虫 ,在培养细胞上清及虫体血淋巴中均测到抗菌活性。经Northernblotting证明感染甜菜夜蛾幼虫中有类CMⅣmRNA的存在。且表达产物在酸性电泳中电泳行为与天然抗菌肽CMⅣ组分相似。为进一步利用昆虫细胞及虫体生产抗菌肽药物打下了基础。     

大肠杆菌海藻糖合成酶基因对提高烟草抗逆性能的研究

编码大肠杆菌海藻糖合成酶的otsA基因由农杆菌介导引入野生型烟草植株并在花椰菜花叶病毒启动子序列 (CaMV35S)控制下获得表达。蒸发光散射高效液相层析法测定海藻糖实验表明 ,转基因烟草能够合成海藻糖 ,合成量达 1 4μg g叶片湿重 ;转基因烟草表现为耐盐性生长、干燥失重缓慢等抗逆表型。说明海藻糖合成酶otsA基因的引入 ,改变了烟草的糖代谢途径 ,同时也提高了植物的耐盐碱、耐干旱特性。     

Glioma models.

Gliomas are primary central nervous system tumors that arise from astrocytes, oligodendrocytes or their precursors. Gliomas can be classified into several groups according to their histologic characteristics, the most malignant of the gliomas is glioblastoma multiforme. In contrast to the long-standing and well-defined histopathology, the underlying molecular and genetic bases for gliomas are only just emerging. Many genetic alterations have been identified in human gliomas, however, establishing unequivocal correlation between these genetic alterations and gliomagenesis requires accurate animal models for this disease. Here we are reviewing the existing animal models for gliomas with different strategies and our current knowledge on the important issues about this disease, such as activation of signal transduction pathways, disruption of cell cycle arrest pathways, cell-of-origin of gliomas, and therapeutic strategies.     

Microbial conversion of n-alkanes into glycolipid biosurfactants, mannosylerythritol lipids, by Pseudozyma (Candida antarctica)

n-Alkanes ranging from C₁₂ to C₁₈ were converted into glycolipid biosurfactants, mannosylerythritol lipids (MEL), by resting cells of Pseudozyma (Candida) antarctica T-34. The highest yield (0.87 g g^–1 substrate) was obtained from 6% (v/v) of n-octadecane after 7 days reaction. The amount of MEL reached 140 g l^–1 by intermittent feeding of the substrate.     

Molecular scaffold of a new pokeweed antifungal peptide deduced by H nuclear magnetic resonance

The antifungal peptide from seeds of Phytolacca Americana (Pokeweed), designated PAFP-S hereinafter, is a recently found cationic peptide which consists of 38 amino acid residues and exihibites a broad spectrum of antifungal activity, including inhibition of certain saprophytic fungi and some plant pathogens. The secondary structure and three cysteine pairings have been investigated by ¹H NMR analysis. The results show that the molecular scaffold of PAFP-S features a triple-stranded antiparallel β-sheet knotted by a typical disulfide bridge motif, which characterizes the knottin fold. CD spectroscopy indicates a high stability of the molecule in solution. Therefore, PAFP-S should be a new member of the knottin structural family and the first antifungal peptide that adopts the knottin-like fold.     

Effects of exogenous hormones on floret development and grain setin wheat

At specific stages during floret development, solutions of IAA,GA₃, zeatin and ABA were injected into the leaf sheath around theyoung spike of wheat (Triticum aestivum L.) to study theregulating effects of exogenous hormones on floret development. Zeatin promotedfloret development and significantly increased the number of fertile florets aswell as grain set, especially at the stage of anther-lobe formation. Zeatinalsoincreased the sugar concentrations in spikes at anthesis. In contrast, IAA,GA₃ and ABA inhibited floret development, with different patternsforeach of the hormones. IAA inhibited the development of the whole spike and allflorets in the spikelets such that grain loss occurred in all positions in thespikelets. GA₃ increased the number of fertile florets per spike,butdecreased grain set of the third floret in each spikelet, especially whenapplied at terminal spikelet formation. ABA inhibited floret development, anddecreased the number of fertile florets and grain set at almost all developmentstages, except at anther-lobe formation. The inhibitory effect of ABA wasmainlyon the first and third florets in each spikelet.     

Organic matter chemistry and dynamics in clear-cut and unmanaged hardwood forest ecosystems

Forest harvesting alters the organic matter cycle by changing litter inputs and the decomposition regime. We hypothesized that these changes would result in differences in organic matter chemistry between clear-cut and uncut watershed ecosystems. We studied the chemistry of soil organic matter (SOM), and dissolved organic carbon (DOC) in soil solutions and stream samples in clear-cut and uncut sites at the Hubbard Brook Experimental Forest in New Hampshire using DOC fractionation techniques and solid-state ¹³C nuclear magnetic resonance (NMR) spectroscopy.Alkyl-C (aliphatic compounds) and O-alkyl-C (carbohydrates) were the largest C fractions in soil and dissolved organic matter at Hubbard Brook. Alkyl-C ranged from 29–48% of soil C, 25–42% of soil solution C, and 22–42% of streamwater DOC. Carbohydrates comprised 32–49%, 36–43%, and 29–60% of C in soils, solutions, and streamwater, respectively. In both soils and soil solutions, the carbohydrate fraction decreased with increasing soil depth, while the aromaticity of organic matter increased with depth. There were no significant differences in the structural chemistry of SOM between clear-cut and uncut watersheds.The aromatic-C fractions in soil solutions at the clear-cut site ranged from 12–16%, approximately 40% greater than at the uncut site (8.5–11%). Thus, clear-cutting has resulted in the leaching of more highly decomposed organic matter, and depletion of more aliphatic compounds in the soluble organic pool. Because DOC fluxes are small compared to the SOM pool, large differences in soil solution chemistry do not substantially alter the overall composition of SOM. While the organic chemistry of stream DOC varied greatly among 3 sampling dates, there were no obvious clear-cutting effects. Thus, temporal variations in flowpaths and/or in-stream processes appear to be more important than disturbance in regulating the organic carbon chemistry of these streams.     

Possible involvement of the mitochondrial alternative pathway in ethylene-induced apoptosis in tomato protoplasts

In this study, a monoclonal antibody to the terminal oxidase of the alternative pathway from Sauromatum guttatum was used to detect the expression of alternative oxidase (AOX) protein in tomato mitochondria. The results show that there was an obvious correlation between the ethylene-induced apoptosis and the levels of AOX protein in tomato cells undergoing ethylene-induced apoptosis. In addition, when tomato protoplasts were preincubated with 2 mM salicylhydroxamic acid (SHAM), an inhibitor of the alternative pathway, before their exposure to ethylene, the TUNEL positive reaction and DNA fragmentation were obviously accelerated. We suggest that AOX may play an important role in protecting tomato protoplasts against ethylene-induced apoptosis.