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21.
用RT-PCR方法快速克隆了Wistar大鼠脑神经元特异性烯醇化酶(NSE)的cDNA,将此包括编码全长NSE433个氨基醚的DNA片段重组入pUC质粒,并用PCR方法测定了全部顺序,经重复实验,发现Wistar大鼠与Forss-Petter报导的SD大鼠NSE基因顺序,有两外单碱基的差别,其中一个涉及氨基酸的改变。同时还对RNA的提取及长片段DNA的RT-PCR扩增进行了方法学的探讨。 相似文献
22.
Membrane association of functional vesicular stomatitis virus matrix protein in vivo. 总被引:26,自引:19,他引:7 下载免费PDF全文
The matrix (M) protein of vesicular stomatitis virus (VSV) is a major structural component of the virion which is generally believed to bridge between the membrane envelope and the ribonucleocapsid (RNP) core. To investigate the interaction of M protein with cellular membranes in the absence of other VSV proteins, we examined its distribution by subcellular fractionation after expression in HeLa cells. Approximately 90% of M protein, expressed without other viral proteins, was soluble, whereas the remaining 10% was tightly associated with membranes. A similar distribution in VSV-infected cells has been observed previously. Conditions known to release peripherally associated membrane proteins did not detach M protein from isolated membranes. Membrane-associated M protein was soluble in the detergent Triton X-114, whereas soluble M protein was not, suggesting a chemical or conformational difference between the two forms. Membranes containing associated M protein were able to bind RNP cores, whereas membranes lacking M protein were not. We suggest that this membrane-bound M fraction constitutes a functional subset of M protein molecules required for the attachment of RNP cores to membranes during normal virus budding. 相似文献
23.
用Northern blot方法对二乙基亚硝胺所诱发的大鼠肝癌中内源性蛋白酶抑制因子α_2-巨球蛋白(α_2-M)、非特异性免疫抑制剂α_1-酸性糖蛋白(α_1-AGP)及雄性激素正调控的α-2u球蛋白(α-2u)三种分泌性蛋白基因表达情况进行了分析。结果表明在大部分(14/16)肝癌样品中α_2-M RNA水平显著降低;而α_1-AGP RNA水平显著高于正常对照水平;α-2u RNA水平明显下降,但在某些雄性大鼠肝癌样品中该基因却有一定程度的表达。这些结果说明,一些肿瘤宿主血浆中α_2-M水平的显著下降及α_1-AGP水平的明显升高分别是由于基因表达活性的下降及升高所致。α-2u基因表达的异常提示,在癌变过程中机体的内分泌功能发生了某些变化。 相似文献
24.
25.
该研究在人工控制水分条件下,设置3个干旱胁迫处理,选用3个主栽油菜品种‘陇油10号’、‘陇油2号’、‘青杂5号’幼苗进行盆栽试验,测定干旱胁迫条件下叶片相对含水量、叶绿素含量、光合气体交换参数及叶绿素荧光参数等指标,考察各指标在干旱胁迫过程中的变化特征,并通过主成分分析(PCA)和隶属函数法评价品种的抗旱性及其主要响应因子,以揭示西北地区油菜幼苗响应干旱胁迫的光合调控机制。结果表明:(1)各品种油菜幼苗的叶片相对含水量(RWC)均随干旱胁迫程度的递增而逐渐降低,最大水分亏缺(WSD)却逐渐上升。(2)各品种油菜幼苗叶片的叶绿素a含量、叶绿素总含量随着干旱胁迫程度的递增而先增加后递减,且同一种幼苗在不同处理间差异显著。(3)各品种油菜幼苗叶片的净光合速率(Pn)、水分利用效率(WUE)、单株生物量、气孔导度(Gs)、胞间CO2浓度(Ci)均在受到干旱胁迫时迅速降低,且同一品种幼苗在不同处理间差异显著,而其叶片蒸腾速率(Tr)在干旱胁迫下无显著变化。(4)各品种油菜幼苗叶片光化学猝灭系数(qP)和非光化学猝灭系数(NPQ)随着干旱胁迫程度的递增先增加后递减,最大光化学效率(Fv/Fm)和电子传递速率(ETR)在受到干旱胁迫时迅速降低,且同一品种幼苗在不同处理间差异显著。(5)主成分分析结果表明,在油菜幼苗受到干旱胁迫时RWC、Ci、Gs、Pn、WUE、叶绿素总含量、叶绿素a含量和NPQ起主要调控作用;隶属函数法综合评价表明,3个品种油菜幼苗耐旱能力由高到低依次为‘陇油10号’>‘陇油2号’>‘青杂5号’。 相似文献
26.
我们用DFY-I型超声图像定量分析诊断仪,测定组织结构超声图像的分贝,灰阶值,本文检测分析了39例正常台盘测值。 相似文献
27.
A biosorbent prepared from Ascophyllum nodosum seaweed biomass, FCAN2, was examined for its sorption capacity. Equilibrium batch sorption studies were performed using two-matal systems containing either (Cu + Zn), (Cu + Cd), or (Zn + Cd). In the evaluation of the two-metal sorption system performance, simple isotherm curves had to be replaced by three-dimensional sorption isotherm surfaces. In order to describe the isotherm surfaces mathematically, three Langmuir-type models were evaluated. The apparent one-parameter Langmuir constant (b) was used to quantify FCAN2 "affinity" for one metal in the presence of another one. The uptake of Zn decreased drastically when Cu or Cd were present. The uptake of Cd wasmuch more sensitive to the presence of Cu than to that of Zn. The presence of Cd and Zn alter the "affinity" of FCAN2 for Cu the least at high Cu equilibrium concentrations. The mathematical model of the two-metal sorption system enabled quantitative estimation of one-metal (bio)sorption inhibition due to the influence of a second metal. (c) 1995 John Wiley & Sons Inc. 相似文献
28.
Molecular dynamics simulations and energy analysis have been carried out to study the structural mobility and stability of the four -helix bundle motifs. The simulation results as well as the X-ray data show that the atomic RMS fluctuation is larger at the loop region for four representative proteins investigated: methemerythrin, cytochrome b-562, cytochrome c, and bovine somatotropin. The loop-loop, helix-helix, and loop-helix interactions are computed for the unfolded and folded proteins. In the folded and solvated protein structures the loop-helix interaction is stronger than the helix-helix interaction, especially in the electrostatic component. But the stabilization energies of both the loop-helix and the helix-helix interactions relative to the those of an unfolded structure are of the same order of magnitude. The stabilization due to protein-solvent interaction is greater in the helix region than in the loop region. The percentage of hydrophilic solvent accessible area for the four proteins studied was calculated with the method of Eisenberg and McLachlan. The percentage of the hydrophilic area is greater in the loops than in the helices. A Poisson-Boltzmann calculation shows that the potential from the loops acting on a helix is generally more negative than that from other helices. 相似文献
29.
Chong D.K.X. Roberts W. Arakawa T. Illes K. Bagi G. Slattery C.W. Langridge W.H.R. 《Transgenic research》1997,6(4):289-296
A 1177 bp cDNA fragment encoding the human milk protein -casein was introduced into Solanum tuberosum cells under control of the auxin-inducible, bidirectional mannopine synthase mas12) promoters using Agrobacterium tumefaciens-mediated leaf disc transformation methods. Antibiotic-resistant plants were regenerated and transformants selected based on luciferase activity carried by the expression vector containing the human -casein cDNA. The presence of human -casein cDNA in the plant genome was detected by PCR and DNA hybridization experiments. Human -casein mRNA was identified in leaf tissues of transgenic plants by RT-PCR analysis. Human - casein was identified in auxin-induced leaf and tuber tissues of transformed potato plants by immunoprecipitation and immunoblot analysis. Human -casein produced in transgenic plants migrated in polyacrylamide gels as a single band with an approximate molecular mass of 30 kDa. Immunoblot experiments identified approximately 0.01% of the total soluble protein of transgenic potato leaf tissue as -casein. The above experiments demonstrate the expression of human milk - casein as part of an edible food plant. These findings open the way for reconstitution of human milk inedible plants for replacement of bovine milk in baby foods for general improvement of infant nutrition, and for prevention of gastric and intestinal diseases in children 相似文献
30.
R. Chong I. S. Maddox M. R. Johns A. G. Wagenaar 《Applied microbiology and biotechnology》1980,9(4):317-323
Summary Fungi which have been previously shown to hydrolyse glycocholic acid, with liberation of the free bile acid, have now been shown to be similarly capable of hydrolysing glycodeoxycholic acid. Sodium taurocholate, however, is much less susceptible and its hydrolysis has been demonstrated with only one of the selected fungi, Penicillium chrysogenum, growing in a medium containing the conjugate as the sole sulphur source. It is concluded that the nature of the amino acid moiety is important in determining the ease of hydrolysis of bile acid conjugates by whole cells of the fungi under test. 相似文献