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971.
大鼠催乳素基因真核细胞可表达性质粒的构建及应用研究 总被引:4,自引:0,他引:4
735bp的PRLcDNA片段从质粒PRL-SP65#1中回收后,用粘性末端连接法将其重组到真核表达载体pcDNA3上,筛选出正向连接重组体pcDNA3-PRLS和反向连接重组体pcDNA3-PRLAS。将重组体pcDNA3-PRLs和空载体pcDNA3分别转入NIH3T3细胞系,用G418筛选出阳性细胞后与未转染的NIH3T3细胞在加E2和不加E2的情况下,用原位杂交的方法,分别用PRLcDNA探针和原癌基因c-H-rascDNA探针进行检测,未转染的NIH3T3细胞在加E2和不加E2时都几乎无催乳素基因的表达,同样,转入空载体的NIH3T3细胞也无PRL的表达,而转入重组体pcDNA3-PRLS的NIH3T3细胞则有大量的PRL基因的表达,与对照组相比有显著差异(P<0.01)。正常和转入空载体的NIH3T3细胞有一定程度的原癌基因c-H-ras的表达,当分别加入E2和转入重组体pcDNA3-PRLS后,NIH3T3细胞中的c-H-ras基因表达水平都显著升高(P<0.05)。 相似文献
972.
经一系列试验证实百日咳菌大罐培养浓度的下降与培养基中不耐热营养因子不足有关,将培养基的高压灭菌方式改为除菌过滤,保留热不稳定营养因子,可以提高百日咳菌大罐培养浓度;优化酸沉淀条件后,提高了菌苗回收率,从而节省了大量培养基,缩短了生产时间,获得了良好的经济效益。 相似文献
973.
The relationship among transfer and expression of auxin binding protein gene (abp), auxin (NAA)-induced plasmalemma hyperpolarity and sensibility to auxin during protoplast culture was studied by measuring transmembrane potential difference (Em) and culturing the protoplasts of sense and antisenseabp transgenic tobacco. The concentration of NAA inducing the highest degree of hyperpolarity of senseabp transgenic tobacco protoplasts was lower than the control, and in protoplast culture, their sensibility to auxin increased. The concentration of antisenseabp transgenic tobacco protoplasts was higher than the control, and in protoplast culture, their sensibility to auxin decreased. These results demonstrated that ABP synthesized in endoplasmic reticulum needed to transport to cell membrane and functioned there. 相似文献
974.
Merete Bjørgan Schrøder Emilio Flaño Lars Pilström Trond Ø. Jørgensen 《Fish & shellfish immunology》1998,8(8):565-576
Localisation of immunoglobulin heavy chain (IgH) producing cells was determined in sections from head kidney, spleen, thymus, gills, gut, skin, heart and liver from the Atlantic cod. In general, IgH mRNA positive cells were detected in all organs examined and were mainly located to the connective tissue surrounding the vascular system in these organs. In the head kidney and spleen, IgH mRNA positive cells appeared as single distributed cells or as dense clusters, whereas in the thymus only single distributed positive cells were observed. The percentage of Ig heavy chain mRNA positive (plasma) cells in the head kidney, spleen and thymus was estimated at about 1% of the total cell mass. The number of IgH mRNA positive cells was lower than this in all the other organs examined. 相似文献
975.
Isabella Di Lernia Alessandra Morana Antonio Ottombrino Stefania Fusco Mosè Rossi M. De Rosa 《Extremophiles : life under extreme conditions》1998,2(4):409-416
Enzymes that convert starch and dextrins to α,α-trehalose and glucose were found in cell homogenates of the hyperthermophilic
acidophilic archaeon Sulfolobus shibatae DMS 5389. Three enzymes were purified and characterized. The first, the S. shibatae trehalosyl dextrin-forming enzyme (SsTDFE), transformed starch and dextrins to the corresponding trehalosyl derivatives with
an intramolecular transglycosylation process that converted the glucosidic linkage at the reducing end from α-1,4 to α-1,1.
The second, the S. shibatae trehalose-forming enzyme (SsTFE), hydrolyzed the α-1,4 linkage adjacent to the α-1,1 bond of trehalosyl dextrins, forming
trehalose and lower molecular weight dextrins. These two enzymes had molecular masses of 80 kDa and 65 kDa, respectively,
and showed the highest activities at pH 4.5. The apparent optimal temperature for activity was 70°C for SsTDFE and 85°C for
SsTFE. The third enzyme identified was an α-glycosidase (SsαGly), which catalyzed the hydrolysis of the α-1,4 glucosidic linkages
in starch and dextrins, releasing glucose in a stepwise manner from the nonreducing end of the polysaccharide chain. The enzyme
had a molecular mass of 313 kDa and showed the highest activity at pH 5.5 and at 85°C.
Received: October 29, 1997 / Accepted: April 29, 1998 相似文献
976.
本研究在初步实现水稻原生质体培养的程序化后,选用普通栽培稻P339和特种稻苏御糯选的原生质体为融合亲本,利用碘乙酸(IA)和罗丹明-6G(R-6G)这两个代谢互补抑制剂钝化处理亲本原生质体,确定了合适的抑制条件。P339用0.25mmol/L IA,苏御糯选用50μg/ml R-6G分别经30min钝化处理,通过PEG和高Ca~(2 )、高pH法诱导融合,异源融合体具有代谢互补效应,经培养得到愈伤组织17块,并进一步分化获得不同形态的再生植株12棵。移栽存活的再生植株成熟后可育,通过对这些植株的形态以及酯酶和过氧化物酶同工酶电泳的分析表明是融合后的体细胞杂种植株。 相似文献
977.
The possibility of enhancing the ex situ bioremediation of a chronically polychlorinated biphenyl (PCB)-contaminated soil
by using Triton X-100 or Quillaya Saponin, a synthetic and a biogenic surfactant, respectively, was studied. The soil, which
contained about 350 mg/kg of PCBs and indigenous aerobic bacteria capable of growing on biphenyl or on monochlorobenzoic acids,
was amended with inorganic nutrients and biphenyl, saturated with water and treated in aerobic batch slurry- and fixed-phase
reactors. Triton X-100 and Quillaya Saponin were added to the reactors at a final concentration of 10 g/l at the 42nd day
of treatment, and at the 43rd and 100th day, respectively. Triton X-100 was not metabolised by the soil microflora and it
exerted inhibitory effects on the indigenous bacteria. Quillaya Saponin, on the contrary, was readily metabolised by the soil
microflora. Under slurry-phase conditions, Triton X-100 negatively influenced the soil bioremediation process by affecting
the availability of the chlorobenzoic acid degrading indigenous bacteria, whereas Quillaya Saponin slightly enhanced the biological
degradation and dechlorination of the soil PCBs. In the fixed-phase reactors, where both the surfactant availability and the
mixing of the soil were lower, Triton X-100 did not exert inhibitory effects on the soil biomass and enhanced significantly
the soil PCB depletion, whereas Quillaya Saponin did not influence the bioremediation process.
Received: 28 April 1998 / Received last revision: 15 July 1998 / Accepted: 29 July 1998 相似文献
978.
Giovanni Savettieri Giacoma Alessandra Mazzola Maria Belén Rodriguez Sanchez Giovanna Caruso Italia Di Liegro Alessandro Cestelli 《Cellular and molecular neurobiology》1998,18(4):369-378
1. Neuronal differentiation depends on crosstalk between genetic program and environmental cues. In this study we tried to dissect this complex interplay by culturing neurons from fetal rat brain cortices in a chemically defined, neuron-specific, medium and on different substrata, either artificial (poly-D-lysine) or natural.2. Among the extracellular matrix compounds used in this study, two (collagen I and fibronectin) allowed only a weak attachment of cortical neurons to the substratum, while the others (collagen IV, laminin, and basal lamina from Engelbreth-Holm-Swarm sarcoma) allowed both firm attachment and moderate to extensive neurite outgrowth from neuronal cell bodies.3. By using synapsin I gene expression as a parameter of neuronal differentiation, we found that neurite outgrowth and neuronal differentiation are not linearly linked. Synapsin I gene expression, in fact, was maximal in neurons cultured on laminin, while the fastest neuritic outgrowth was recorded in cultures on poly-D-lysine.4. The data presented in this paper are consistent with the hypothesis that the extracellular matrix plays an active role in modulating the differentiative program of neurons. 相似文献
979.
Luciana Di Ciero Maria L. V. Oliva Ricardo Torquato Peter Köhler Jürgen K. P. Weder José Camillo Novello Claudio A. M. Sampaio Benedito Oliveira Sergio Marangoni 《Journal of Protein Chemistry》1998,17(8):827-834
Trypsin inhibitors of two varieties of Bauhinia variegata seeds have been isolated and characterized. Bauhinia variegata candida trypsin inhibitor (BvcTI) and B. variegata lilac trypsin inhibitor (BvlTI) are proteins with M
r
of about 20,000 without free sulfhydryl groups. Amino acid analysis shows a high content of aspartic acid, glutamic acid, serine, and glycine, and a low content of histidine, tyrosine, methionine, and lysine in both inhibitors. Isoelectric focusing for both varieties detected three isoforms (pI 4.85, 5.00, and 5.15), which were resolved by HPLC procedure. The trypsin inhibitors show K
i
values of 6.9 and 1.2 nM for BvcTI and BvlTI, respectively. The N-terminal sequences of the three trypsin inhibitor isoforms from both varieties of Bauhinia variegata and the complete amino acid sequence of B. variegata var. candida L. trypsin inhibitor isoform 3 (BvcTI-3) are presented. The sequences have been determined by automated Edman degradation of the reduced and carboxymethylated proteins of the peptides resulting from Staphylococcus aureus protease and trypsin digestion. BvcTI-3 is composed of 167 residues and has a calculated molecular mass of 18,529. Homology studies with other trypsin inhibitors show that BvcTI-3 belongs to the Kunitz family. The putative active site encompasses Arg (63)–Ile (64). 相似文献
980.
V Di Marzo 《Life sciences》1999,65(6-7):645-655
The two putative endogenous ligands of cannabinoid receptors, anandamide and 2-arachidonoylglycerol, are synthesized by and released from neurons in a Ca2+-dependent fashion, and re-uptaken and catabolized by both neurons and astrocytes. These biochemical features of the endocannabinoids, as well as some of their pharmacological effects in both central and peripheral nervous systems, suggest a role as neuromodulators for these metabolites. This neuromodulatory role is supported by the brain regional distribution of anandamide, its biosynthetic precursor and its major inactivating enzyme, and by the existence of possible regulatory mechanisms for the biosynthesis and inactivation of endocannabinoids, which are reviewed in this article. 相似文献