Structural and Mutational Studies on Substrate Specificity and Catalysis of Salmonella typhimurium D-Cysteine Desulfhydrase

Salmonella typhimurium DCyD (StDCyD) is a fold type II pyridoxal 5' phosphate (PLP)-dependent enzyme that catalyzes the degradation of D-Cys to H2S and pyruvate. It also efficiently degrades beta-chloro-D-alanine (beta CDA). D-Ser is a poor substrate while the enzyme is inactive with respect to L-Ser and 1-amino-1-carboxy cyclopropane (ACC). Here, we report the X-ray crystal structures of StDCyD and of crystals obtained in the presence of D-Cys, beta CDA, ACC, D-Ser, L-Ser, D-cycloserine (DCS) and L-cycloserine (LCS) at resolutions ranging from 1.7 to 2.6 angstrom. The polypeptide fold of StDCyD consisting of a small domain (residues 48-161) and a large domain (residues 1-47 and 162-328) resembles other fold type II PLP dependent enzymes. The structures obtained in the presence of D-Cys and beta CDA show the product, pyruvate, bound at a site 4.0-6.0 angstrom away from the active site. ACC forms an external aldimine complex while D- and L-Ser bind non-covalently suggesting that the reaction with these ligands is arrested at C alpha proton abstraction and transimination steps, respectively. In the active site of StDCyD cocrystallized with DCS or LCS, electron density for a pyridoxamine phosphate (PMP) was observed. Crystals soaked in cocktail containing these ligands show density for PLP-cycloserine. Spectroscopic observations also suggest formation of PMP by the hydrolysis of cycloserines. Mutational studies suggest that Ser78 and Gln77 are key determinants of enzyme specificity and the phenolate of Tyr287 is responsible for C alpha proton abstraction from D-Cys. Based on these studies, a probable mechanism for the degradation of D-Cys by StDCyD is proposed.

Photocytotoxic ferrocene-appended (L-tyrosine)copper(II) complexes of phenanthroline bases

Copper(II) complexes of ferrocene(Fc)-conjugated reduced Schiff base of L-tyrosine (Fc-TyrH), viz., Cu(Fc-Tyr)(L)](ClO4), where L is 1,10-phenanthroline (phen, 1), dipyrido3,2-d:2',3'-f]quinoxaline (dpq, 2), dipyrido3,2-a:2',3'-c]phenazine (dppz, 3) and 2-(naphthalen-1-yl)-1H-imidazo4,5-f]1,10]phenanthroline (nip, 4), were prepared and tested for their photocytotoxicity in cancer cells. Cu(Fc-Phe)(phen)](-ClO4) (5) of L-phenylalanine and Cu(Ph-Tyr)(L)(ClO4)] of the reduced Schiff base Ph-TyrH derived from benzaldehyde and L-tyrosine having phen (6) and dppz (7), and Cu(Ph-Phe)(phen)(ClO4)] (8) using L-phenylalanine were prepared and used as controls. Complexes 5 and 6 were structurally characterized by X-ray crystallography. A copper(II)-based d-d band near 600 nm and a ferrocenyl band at similar to 450 nm were observed in DMF-Tris-HCI buffer (1:4 v/v) in respective complexes. The complexes are photocleavers of pUC19 DNA in visible light forming (OH)-O-center dot radicals. They are cytotoxic in HeLa (human cervical cancer) and MCF-7 (human breast cancer) cells showing an enhancement of cytotoxicity in visible light. Fluorescence imaging shows nuclear localization of the complexes.

Effect of hybridizing micron-sized Ti with nano-sized SiC on the microstructural evolution and mechanical response of Mg-5.6Ti composite

In this work, the effect of hybridizing micro-Ti with nano-SiC particulates on the microstructural and the mechanical behaviour of Mg-5.6Ti composite were investigated. Mg materials containing micron-sized Ti particulates hybridized with different amounts of nano-size SiC particulates were synthesized using the disintegrated melt deposition method followed by hot extrusion. The microstructural and mechanical behaviour of the developed Mg hybrid composites were studied in comparison with Mg-5.6Ti. Microstructural characterization revealed grain refinement attributed to the presence of uniformly distributed micro-Ti particles embedded with nano-SiC particulates. Electron back scattered diffraction (EBSD) analyses of Mg-(5.6Ti + 1.0SiC)(BM) hybrid composite showed relatively more localized recrystallized grains and lesser tensile twin fraction, when compared to Mg-5.6Ti. The evaluation of mechanical properties indicated that the best combination of strength and ductility was observed in the Mg-(5.6Ti + 1.0SiC)(BM) hybrid composites. The superior strength properties of the Mg-(5.6Ti + x-SiC)(BM) hybrid composites when compared to Mg-5.6Ti is attributed to the presence of nano-reinforcements, the uniform distribution of the hybridized particles and the better interfacial bonding between the matrix and the reinforcement particles, achieved by nano-SiC addition.

Molecular Dynamics Perspective on the Protein Thermal Stability: A Case Study Using SAICAR Synthetase

The enzyme SAICAR synthetase ligates aspartate with CAIR (5'-phosphoribosyl-4-carboxy-5-aminoimidazole) forming SAICAR (5-amino-4-imidazole-N-succinocarboxamide ribonucleotide) in the presence of ATP. In continuation with our previous study on the thermostability of this enzyme in hyper-/thermophiles based on the structural aspects, here, we present the dynamic aspects that differentiate the mesophilic (E. coli, E. chaffeensis), thermophilic (G. kaustophilus), and hyperthermophilic (M. jannaschii, P. horikoshii) SAICAR synthetases by carrying out a total of 11 simulations. The five functional dimers from the above organisms were simulated using molecular dynamics for a period of 50 ns each at 300 K, 363 K, and an additional simulation at 333 K for the thermophilic protein. The basic features like root-mean-square deviations, root-mean-square fluctuations, surface accessibility, and radius of gyration revealed the instability of mesophiles at 363 K. Mean square displacements establish the reduced flexibility of hyper-/thermophiles at all temperatures. At the simulations time scale considered here, the long-distance networks are considerably affected in mesophilic structures at 363 K. In mesophiles, a comparatively higher number of short-lived (having less percent existence time) C alpha, hydrogen bonds, hydrophobic interactions are formed, and long-lived (with higher percentage existence time) contacts are lost. The number of time-averaged salt-bridges is at least 2-fold higher in hyperthermophiles at 363 K. The change in surface accessibility of salt-bridges at 363 K from 300 K is nearly doubled in mesophilic protein compared to proteins from other temperature classes.

Phenyl carbohydrazone conjugated 2-oxoindoline as a new scaffold that augments the DNA and BSA binding affinity and anti-proliferative activity of a 1,10-phenanthroline based copper(II) complex

A new type of copper(II) complex, CuL(phen)(2)](NO3) (CuIP), where L ((E)-N'-(2-oxoindolin-3-ylidene) benzohydrazide) is a N donor ligand and phen is the N, N-donor heterocyclic 1,10-phenanthroline, has been synthesized. The phenyl carbohydrazone conjugated isatin-based ligand L and CuIP were characterized by elemental analysis, infrared, UV-Vis, H-1 and C-13 NMR and ESI-mass spectral data, as well as single-crystal X-ray diffraction. The interaction of calf thymus DNA (CT DNA) with L and CuIP has been investigated by absorption, fluorescence and viscosity titration methods. The complex CuIP displays better binding affinity than the ligand L. The observed DNA binding constant (K-b = 4.15(+/- 0.18) x 10(5) M-1) and binding site size (s = 0.19), viscosity data together with molecular docking studies of CuIP suggest groove binding and/or a partial intercalative mode of binding to CT DNA. In addition, CuIP shows good binding propensity to the bovine serum albumin (BSA) protein, giving a K-BSA value of 1.25(+/- 0.24) x 10(6) M-1. In addition, the docking studies on DNA and human serum albumin (HSA) CuIP interactions are consistent with the consequence of binding experiments. The in vitro anti-proliferative study establishes the anticancer potency of the CuIP against the human cervical (HeLa) and breast (MCF7) cancer cells; noncancer breast epithelial (MCF10a) cells have also been investigated. CuIP shows better cytotoxicity and sensitivity towards cancer cells over noncancer ones than L under identical conditions, with the appearance of apoptotic bodies. (C) 2014 Elsevier B.V. All rights reserved.

Thermal Decomposition of 1,2-Dichloroethane in a Single-Pulse Shock Tube

在单脉冲激波管上,研究了1,2-二氯乙烷的热裂解.实验的激波条件为:温度区间1020 K＜T＜1190 K, 压力: P=0.12 MPa,实验时间τ=0.5 ms;实验气体为1,2-二氯乙烷稀释于Ar气中(3.95 mmol/L).以4-甲基-1-环己烯作为对比速率法实验的内标物,用4-甲基-1-环己烯开环反应的速率常数k=1015.3exp(-33400/T) s-1,以及从其产物的浓度推定出实验温度.经激波加热后的实验气体的终产物用气相色谱分析出主要成分为C2H3Cl,指示出主要反应通道为β消去反应.如把所有产物C2H3Cl都归于β消去反应,则可推定出表观之反应速率常数k1a=5.0×1013exp(-30000/T) s-1.对于由C-Cl键断键反应引发的链反应的可能影响做了分析研究.用了一种简便分析可推知在实验的温度范围内的低端(1020 K)链反应的影响可以忽略,而在其高端(1190 K)链反应将给出10%的终产物C2H3Cl的附加浓度,获得真实的β消去反应速率常数则必须把这部分予以扣除.经过这样的校正之后,最后得到CH2ClCH2Clβ消去反应速率常数为k1c=2.3×1013exp(-29200/T) s-1.

High performance nanocomposite thin film transistors with bilayer carbon nanotube-polythiophene active channel by ink-jet printing

Nanocomposite thin film transistors (TFTs) based on nonpercolating networks of single-walled carbon nanotubes (CNTs) and polythiophene semiconductor [poly [5, 5′ -bis(3-dodecyl-2-thienyl)- 2, 2′ -bithiophene] (PQT-12)] thin film hosts are demonstrated by ink-jet printing. A systematic study on the effect of CNT loading on the transistor performance and channel morphology is conducted. With an appropriate loading of CNTs into the active channel, ink-jet printed composite transistors show an effective hole mobility of 0.23 cm 2 V-1 s-1, which is an enhancement of more than a factor of 7 over ink-jet printed pristine PQT-12 TFTs. In addition, these devices display reasonable on/off current ratio of 105-10 6, low off currents of the order of 10 pA, and a sharp subthreshold slope (<0.8 V dec-1). The work presented here furthers our understanding of the interaction between polythiophene polymers and nonpercolating CNTs, where the CNT density in the bilayer structure substantially influences the morphology and transistor performance of polythiophene. Therefore, optimized loading of ink-jet printed CNTs is crucial to achieve device performance enhancement. High performance ink-jet printed nanocomposite TFTs can present a promising alternative to organic TFTs in printed electronic applications, including displays, sensors, radio-frequency identification (RFID) tags, and disposable electronics. © 2009 American Institute of Physics.

Evaluación de dosis del herbicida Pyribenxozim (Pyanchor 5 ec) aplicados en post emergencia temprana, para el control de arvenses en época seca en el cultivo de arroz (Oryza sativa L) Timal, Tipitapa, 2006

El presente estudio se realizó en febr ero del año 2006, en Tipi tapa, municipio del departamento de Managua, Km 26 carretera norte. En la zona arrocera el Timal, finca del pivote 34, del señor Freddy González, con el objetivo de ev aluar la eficiencia de tres dosis del herbicida Pyribenxozim (pyanc hor 5 EC) aplicado en post emergencia temprana para el control de arvenses en el cultivo de arroz (Oryza sativa L), de riego en época seca. El experimento se estableció en un lote comercial, en un Diseño de Bloque Completo al Azar (BCA), con cinco tratamientos y cuatro repeticiones. Las variables evaluadas en este estudio fueron: composición florística, cobertura, Efectividad del herbicida sobre las arvenses selectividad (Fitotoxicidad) y rendimiento. Las dosis del producto evaluado en el ensayo fueron: T1- (Pyribenxozim 5 EC 0.8 l/ha), T2 - (Pyribenxozim 5 EC 1.0 l/ha), T3 - (Pyribenxozim 5 EC 1.2 l/ha), T4 - (Testigo comercial Bispiribac Sodico (Nominee) 40 SC 1.0 l/ha) (muy utilizado en la zona) y T5 - (Testigo absoluto). En los resultados, se identificaron las arvenses que predominaban en el cultivo, las principales familias predominantes fueron las Poaceae. Los mayores porcentajes de c obertura se encontraron en T1- (Pyribenxozim 5 EC 0.8 l/ha) y el T5 - (Testigo absoluto). Los tres tratamientos evaluados de pyanchor 5 EC controlaron eficientemente las principales arvenses presentes. Sin embargo el mejor resultado sobre el detrimento de las arvenses fue la dosis de 1.2 l/ ha de Pyanchor 5 EC en la que se encontró un 97 % de control. En cuanto a la efectividad sobre el tipo de especies, se observó, que ninguna de las dosis estudiadas logro afectar a las especies Leptochloa filiformis L y Eclipta alba L mismo efecto se observó en el tratamiento en el que se uso el producto comercial Bispir ibac Sodico (Nominee) 40 SC 1.0 l/ha. El herbicida Pyribenxozim (pyanchor 5 EC), es un producto selectivo con respecto al cultivo de arroz. Según esto s resultados, en el rendimien to del cultivo ninguno de los tratamientos afecto su calidad y cantidad según características de la variedad. En cuanto a la probación y registro del producto por el MAG FOR, basado en los resultados obtenidos este actualmente fue aceptado y esta siendo aplicado por los productores de arroz.

Prevalence Of Shear Banding In Compression Of Zr41Ti14Cu12.5Ni10Be22.5 Pillars As Small As 150 Nm In Diameter

Recently, the size dependence of mechanical behaviors, particularly the yield strength and plastic deformation mode, of bulk metallic glasses (BMG) has created a great deal of interest. Contradicting conclusions have been drawn by different research groups, based on various experiments on different BMG systems. Based on in situ compression transmission electron microscopy (TEM) experiments on Zr41Ti14Cu12.5Ni10Be22.5 (Vit 1) nanopillars, this paper provides strong evidence that shear banding still prevails at specimen length scales as small as 150 nm in diameter. This is supported by in situ and ex situ images of shear bands, and by the carefully recorded displacement bursts under load control its well as load drops under displacement control. Finite element modeling of the stress state within the pillar shows that the unavoidable geometry constraints accompanying such experiments impart a strong effect on the experimental results, including non-uniform stress distributions and high level hydrostatic pressures. The seemingly improved compressive ductility is believed to be due to such geometry constraints. Observations underscore the notion that the mechanical behavior of metallic glasses, including strength and plastic deformation mode, is size independent at least in Vit 1. (C) 2009 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Feeding and waste management strategies in the culture of Clarias gariepinus

Nutrient digestibility and amino acid availability were assessed in sharp-toothed catfish, Clarias gariepinus, fingerlings fed diets containing soyabean flour (SF) - Poultry meat meal (PMM) blends (25:75. 50:50, and 75:25) and 0.5 of 1.0%, Cr sub(2)0 sub(3). There was agreement between the pattern of overall protein digestibility and average amino acid availability despite the variability in individual amino acid availability the best dry matter, lipid and protein digestibility coefficients, and amino acid availability values were obtained with diets containing 0.5% Cr sub(2)0 sub(3). Chromic Oxide inclusion level appeared to affect nutrient availability. Increased marker level resulted into decreased nutrient digestibility coefficients. Similarly, these diets generated lower fecal crude protein than those with 1.0% Cr sub(2)0 sub(3). However, the latter group recorded higher protein retention efficiency. Dry mailer and lipid of diets containing more soyabean flour seemed to be more digestible than those of poultry meat meal. Similar trend was observed for the apparent availability of the amino acids. This investigation has indicated that low level of marker was better in digestibility study. Utilization of more SF than PMM in the diets of this catfish was more beneficial and should be encouraged in the feed industries producing catfish diets towards a better feed and waste management strategies in this aquaculture operation

Avaliação das propriedades de barreira a gases de membranas obtidas a partir de dispersões aquosas à base de poliuretanos e argila

Materiais nanoestruturados têm recebido destaque na comunidade científica, destacando-se, dentre eles, os nanocompósitos à base de polímeros e argila. Quando esses materiais são obtidos no estado líquido, ressalta-se também o uso de água em substituição a solventes orgânicos, devido a questões ambientais. Neste trabalho foram sintetizadas dispersões aquosas à base de poliuretanos (WPUs) e argila hidrofílica do tipo montimorilonita (MMT) de natureza sódica, com o objetivo de avaliar as propriedades de barreira a gases conferidas pela presença de argila e pela variação nas proporções entre os segmentos flexíveis poli(glicol propilênico) (PPG) e o copolímero em bloco à base de poli(glicol etilênico) e poli(glicol propilênico) (EG-b-PG). Os monômeros empregados na síntese foram: poli(glicol propilênico) (PPG); copolímero em bloco à base de poli(glicol etilênico) e poli(glicol propilênico) (EG-b-PG), com teor de 7% de EG; ácido dimetilolpropiônico (DMPA), diisocianato de isoforona (IPDI) e etilenodiamina (EDA), como extensor de cadeia. Foram sintetizadas dispersões aquosas com e sem a presença de argila, fixando-se a razão entre o número de equivalentes-grama de grupos diisocianato e hidroxila (razão NCO/OH) em 1,5. Nas formulações foi variado também o teor de argila em relação à massa de prepolímero em 0,5% e 1%. Foi adicionada uma etapa de agitação adicional com dispersor Turrax em algumas formulações. A argila foi previamente deslaminada em água deionizada e incorporada à formulação na etapa da dispersão do prepolímero. As dispersões foram avaliadas, quanto ao teor de sólidos totais, tamanho médio de partícula e viscosidade aparente. Os filmes vazados a partir das dispersões foram caracterizados por espectrometria na região do infravermelho (FTIR) e permeabilidade ao CO2. A resistência térmica dos filmes foi determinada por termogravimetria (TG). Foram observadas modificações nas propriedades dos filmes obtidos com a inserção da argila e com a variação no teor de segmentos à base de poli(glicol etilênico). A inserção da argila promoveu uma melhoria na resistência térmica das membranas bem como uma redução na permeabilidade das mesmas. Foi observado um aumento na permeabilidade das membranas obtidas a partir das formulações com maior percentual de copolímero (EG-b-PG), com e sem argila.

Influence of Extracellular Matrix Components on the Expression of Integrins and Regeneration of Adult Retinal Ganglion Cells

Purpose Retinal ganglion cells (RGCs) are exposed to injury in a variety of optic nerve diseases including glaucoma. However, not all cells respond in the same way to damage and the capacity of individual RGCs to survive or regenerate is variable. In order to elucidate factors that may be important for RGC survival and regeneration we have focussed on the extracellular matrix (ECM) and RGC integrin expression. Our specific questions were: (1) Do adult RGCs express particular sets of integrins in vitro and in vivo? (2) Can the nature of the ECM influence the expression of different integrins? (3) Can the nature of the ECM affect the survival of the cells and the length or branching complexity of their neurites? Methods Primary RGC cultures from adult rat retina were placed on glass coverslips treated with different substrates: Poly-L-Lysine (PL), or PL plus laminin (L), collagen I (CI), collagen IV (CIV) or fibronectin (F). After 10 days in culture, we performed double immunostaining with an antibody against beta III-Tubulin to identify the RGCs, and antibodies against the integrin subunits: alpha V, alpha 1, alpha 3, alpha 5, beta 1 or beta 3. The number of adhering and surviving cells, the number and length of the neurites and the expression of the integrin subunits on the different substrates were analysed. Results PL and L were associated with the greatest survival of RGCs while CI provided the least favourable conditions. The type of substrate affected the number and length of neurites. L stimulated the longest growth. We found at least three different types of RGCs in terms of their capacity to regenerate and extend neurites. The different combinations of integrins expressed by the cells growing on different substrata suggest that RGCs expressed predominantly alpha 1 beta 1 or alpha 3 beta 1 on L, alpha 1 beta 1 on CI and CIV, and alpha 5 beta 3 on F. The activity of the integrins was demonstrated by the phosphorylation of focal adhesion kinase (FAK). Conclusions Adult rat RGCs can survive and grow in the presence of different ECM tested. Further studies should be done to elucidate the different molecular characteristics of the RGCs subtypes in order to understand the possible different sensitivity of different RGCs to damage in diseases like glaucoma in which not all RGCs die at the same time.

Avaliação in vitro da incorporação de clorexidina nas propriedades físico-químicas de resinas macias

A incorporação do diacetato de clorexidina em resinas macias para reembasamento de próteses pode se tornar uma boa alternativa na terapia com antimicrobianos usados para candidíase oral, uma vez que é independente da cooperação do paciente. Este trabalho teve como objetivo analisar a hipótese que a incorporação de diacetato de clorexidina em resinas macias não alteraria as suas propriedades físico-químicas. Foram analisadas duas resinas macias, uma à base de polimetilmetacrilato (PMMA) e outra à base de polietilmetacrilato (PEMA), com a incorporação de 0,5%, 1,0% e 2,0% de diacetato de clorexidina, além do grupo controle sem diacetato de clorexidina. Os espécimes foram armazenadas em água destilada a 37 durante 2hrs, 48 hrs, 7, 14, 21 e 28 dias e analisados quanto a cristalinidade do polímero através da difração de Raios X, a dureza mensurada com durômetro na escala shore A, o grau de conversão dos monômeros pela Espectroscopia de Infravermelho Transformada de Fourier (FTIR), a lixiviação de monômeros residuais e a liberação da clorexidina por Cromatografia Líquida de Alta Performace (HPLC). Os resultados mostraram que quanto à cristalinidade dos polímeros nas resinas macias à base de PMMA e de PEMA não alteraram após a incorporação de clorexidina, o que está relacionado à dureza Shore A, que aumentou ao longo do tempo, mas sem alteração significativa para nenhuma das concentrações de diacetato de clorexidina testadas. Já quanto ao grau de conversão, após a incorporação de clorexidina, a resina macia à base de PMMA não apresentou diferença estatisticamente significativa (р>0,05), porém a resina macia à base de PEMA apresentou diminuição significativa do grau de conversão (p<0,05), o que refletiu no aumento significativo da lixiviação de monômeros residuais nas concentrações 0,5% e 1,0% de clorexidina (p<0,05), principalmente nas primeiras 48 horas nessa resina. A resina macia à base de PMMA não apresentou alterações do padrão de lixiviação desses monômeros (р>0,05) e para ambas as resinas a cinética de liberação de clorexidina foi relacionada à lixiviação de monômeros residuais, que para a resina macia à base de PEMA foi maior significativamente nas primeiras 48 horas (p<0,05) e para a resina macia à base de PMMA não teve alteração estatisticamente significativa (р>0,05) se mantendo até o último dia de análise. Desse modo, conclui-se que a incorporação do diacetato de clorexidina não afetou a cristalinidade dos polímeros, inalterando a dureza nas duas resinas, porém o grau de conversão sofreu diminuição após a incorporação de diacetato de clorexidina na resina macia à base de PEMA, o que levou ao aumento da lixiviação de monômeros residuais e curta liberação de clorexidina; enquanto que a resina macia à base de PMMA, devido a não alteração significativa do seu grau de conversão, ocorreu menor lixiviação de monômeros residuais e liberação prolongada de clorexidina.

小麦春化相关基因的克隆及其功能分析

一． 小麦相关基因ver203F cDNA全序列克隆与功能分析 　　根据本实验室通过差异筛选技术克隆到的与春化相关的基因cDNA verc203的序列，设计PCR 5’端PCR引物，利用RACE（rapid amplification of cDNA ends）克隆策略，得到春化相关基因ver203基因的同源基因ver203F cDNA的3’端序列，长度为1,197bp。Northern分析表明ver203F全长约为1.5 kb，且其表达具有春化处理的特异性。根据3’RACE克隆的ver203F 3 ’端核苷酸序列设计了3’端PCR引物，利用5’RACE克隆到该基因的5’端片段，经DNASIS核酸分析软件分析将5’45RACE和3’RACE DNA序列拼接合并，得到ver203F全长cDNA，从TdT加尾5’末端到poly A全长为1,561 bp，5’端起始密码子ATG上游非编码区－1～－192共了192bp，终止密码子TGA到poly A的非编码区有253bp，cDNA编码区全长1,119 bp，推测编码373氨基酸残基。国际基因序列数据库检索表明该基因序列（GenBank/EMBL/DDBJ：AB012013）与大麦茉莉酸诱导基因有部分同源性。因上推测该基因在调控开花过程中可能参与茉莉酸介导的信号传导途径，ver203F作用的发挥可能需要其它蛋白的参与，或ver203F本身就是一个受体蛋白。 　　为了研究ver203F基因的功能，将通过3’RACE克隆到的ver203F 3’端序到分别构建正义和反义植物表达载体，通过花粉管通道法、农杆菌介导的叶圆片法以及农杆菌介导的真空转化法分别转化小麦、烟草和拟南芥菜。获得转基因植株后，PCR、DNA Dob Blot、Southern Blot分析以及GUS活性检测证明外源基因已整合到转基因植株中，并得到表达。在获得的小麦、烟草和拟南芥菜转基因植株中，它们开花时间都相应地推迟，表明正常植物体内该基因在控制营养生长向生殖生长的转变中起作用。ver203F可以影响小麦和拟南茶菜花序的发育，首先无论正义还是反义都使得花序的发育受到抑制，在小麦中表现为顶部小穗退化，在拟南芥菜中表现为顶花。其次在转化正义基因的转基因拟南芥菜中，观察到产生的顶花为对称的两朵或以对称的两朵顶花基部为生长点长出丛生花，这种对称花的麦型小麦小穗中小花的表型相类似，说明ver203F基因可能在小麦小花的发育过程中也起着重要作用。 二． 春化相关基因ver17在开花过程中功能的分析 以春化处理冬小麦（京冬1号）幼苗cDNA为材料，通过减法杂交与差异筛选得到春化相关cDNA克隆verc17。为了研究该基因的功能，以包含CaMV 35S启动子的pBI221为载体，将ver17cDNA片段分别从两个方向插入pBI221的BamH I－Sma I, Xba I－BamH I间，构建正义和反义表达质粒：p17S和p17X，通过花粉管通道法转化小麦。对T0和T1两代转基因小麦的观察发现，在转化反义基因p17X的转基因小麦首先表现为抽穗推迟，其次穗的顶部和基部小穗严重退化，另外还发现转化反义基因的小麦败育现象严重（主要是花粉败育），因此推测ver17基困能可具有以下几方面的特点：a．春化诱导型表达;b．促进植物开花;c．促进穗顶端和基部花的发育，减少其退化;d．影响雄蕊的发育。