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991.
We have synthesised the -subunit of the chick nicotinic acetylcholine receptor (nAChR) in stable, continuous insect (Spodoptera frugiperda) cell lines. A cDNA was integrated randomly into the insect cell genome under control of a baculovius immediate early gene promoter. Transformed cells were obtained by co-transfection of the insect cells with pIEK1.nAChR, encoding the -subunit cDNA, and pIEK1.neo, encoding the neomycin resistance gene. G-418-resistant clones were selected and expanded into continuous cell lines synthesising the chick nAChR -subunit. Using fluorescence microscopy and ligand binding studies we were able to demonstrate efficient membrane targeting of the receptor subunit in the insect cell plasma membrane. Stable insect cell lines may thus have significant advantages over transient baculovirus vectors for the synthesis and characterisation of heterologous receptor proteins.Abbreviations AcNPV
Autographa californica nuclear polyhedrosis virus
- BTX
-bungarotoxin
- BSA
bovine serum albumin
- FITC
Fluoroscein isothiocyanate
- G418
geneticin-418
- hpi
hours post-infection
-
ie-1
immediate early 1 gene
- nAChR
nicotinic acetylcholine receptor alpha subunit
- Sf
Spodoptera frugiperda
- tPA
tissue plasminogen activator 相似文献
992.
Shinji Hosoi Mitsuo Satoh Hiromasa Miyaji Tatsunari Nishi Tamio Mizukami Mamoru Hasegawa Seiga Itoh Tatsuya tamaoki 《Cytotechnology》1995,19(1):1-10
Pro-UKS1 was designed as a thrombin-resistant derivative of pro-urokinase (pro-UK) by introducing a glycosylation site using site-directed mutagenesis. An expression plasmid for pro-UKS1, pMo1UKS1SEd1-5, was constructed and introduced into Namalwa KJM-1 cells (Hosoiet al., 1988), and cells resistant to G418 and Methotrexate (MTX) were obtained. Amongst them, the highest pro-UKS1 producer (resistant to 500 nM of MTX), clone 41-8, was selected and further characterized. Clone 41-8 was cultured in serum-free ITPSGF medium (Hosoiet al., 1988). Under the conventional conditions, the concentration of pro-UKS1 reached 26 g ml–1. Addition of glucose and tri-iodothyronine (T3) improved productivity, and the maximal productivity of pro-UKS1 was 67 g ml–1 day–1. In this conditioned medium, content of pro-UKS1 was above 80% of total proteins.Abbreviations BSA
bovine serum albumin
- dhfr
dihydrofolate reductase
- HEPES
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- kb
kilobase pairs
- kDa
kilodaltons
- MTX
Methotrexate
- PBS
phosphate buffered saline
- pro-UK
pro-urokinase
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- T3
tri-iodothyronine
- Tween-PBS
phosphate buffered saline containing 0.05% Tween 80 相似文献
993.
V. Cusumano G.B. Costa R. Trifiletti R.A. Merendino G. Mancuso 《FEMS immunology and medical microbiology》1995,10(2):151-156
Abstract Contamination of food with mycotoxins is a major health problem. Impairment of several immune functions has been repeatedly reported in animals fed with contaminated fodder. Since the liver is a major target of toxicity by aflatoxins, the effects of aflatoxins B1, and its hepatic metabolites Q1 and M1 on Kupffer cell function was investigated in vitro. Aflatoxin B1 induced significant ( P < 0.05) inhibition of phagocytosis, intracellular killing of Candida albicans , and intrinsic anti-Herpes virus activity at concentrations as low as 0.01 pg ml−1 . Aflatoxin Q1 and M1 had similar effects on phagocytosis and microbicidal activity, but were two- to ten-fold less potent than aflatoxin B1 . 相似文献
994.
Summary Aside from its role as one of the limiting essential amino acids in protein metabolism, tryptophan (TRP) serves as precursor for the synthesis of the neurotransmitters serotonin and tryptamine as well as for the synthesis of the antipellagra vitamin nicotinic acid and the epiphyseal hormone melatonin.By involvement in so manifold pathways, TRP and its metabolites regulate neurobehavioral effects such as appetite, sleeping-waking-rhythm and pain perception. TRP is the only amino acid which binds to serum albumin to a high degree. Its transport through cell membranes is competetrvely inhibited by large neutral amino acids (NAA). The TRP/NAA ratio in plasma is essential for the TRP availability and thus for the serotonin synthesis in the brain.Due to its high TRP-concentration, human milk protein provides optimal conditions for the availability of the neurotransmitter serotonin. Low protein cow's milk-based infant formulas supplemented with-lactalbumin — a whey protein fraction containing 5.8% TRP — present themselves as a new generation of formulas, with an amino acid pattern different from the currently used protein mixtures of adapted formulas, resembling that of human milk to a much higher degree. 相似文献
995.
Krystyna Harmata 《Vegetation History and Archaeobotany》1995,4(4):235-243
The pollen diagram from Tarnowiec concentrates on human impact on vegetation, and is supported by the archaeological background. The earliest evidence of human activities dated at ca. 6500 B.P. was probably connected with the Neolithic Linearbandkeramik culture. Further periods of settlement are recorded at ca. 5200 B.P. and at 4800 B.P. The next very distinct period of intensification in human impact at ca. 4200-3800 B.P. was most certainly connected with the presence of the Corded Ware culture. The next period of settlement occurred during the late Bronze Age, in connection with the development of the Lusatian culture from ca. 3200 B.P. but is recorded only faintly in the pollen diagram. The next colonization phase at ca. 2600 B.P. is possibly the result of Scythian invasions. The widest expansion of human impact came during the period of Roman influence. After this, anthropogenic indicators decreased during the Migration period. The youngest part of the pollen diagram reveals an intensification of anthropogenic indicators connected with early Medieval settlement. 相似文献
996.
In the fowl the primordial germ cells accumulate in the germinal crescent to the anterior of the two-day embryo. A simple ballistic device has been used to fire tungsten particles (mean diameter 1.5 m) into this region. By coating these projectiles with vector DNA it is possible to transfect these cells. Hatchlings produced by this technique were raised to sexual maturity and shown to contain the foreign DNA in their sperm. G1 offspring containing this DNA were also produced in roughly 20% of these cockerels. In the majority of cases the vector DNA disappeared from the G1 generation as they matured suggesting that in these cases it had been transmitted episomally. 相似文献
997.
内皮素-1对缺氧肺动脉平滑肌细胞的增殖作用 总被引:2,自引:0,他引:2
内皮素(ET)是至今所发现的最强的内源性血管收缩肽,近年来发现ET-1能促进血管平滑肌细胞增殖。本研究表明ET-1对缺氧培养的肺动脉平滑肌细胞(PASMC)有剂量依赖的增殖作用,缺氧可促进PASMC的DNA合成且增加ET-1的丝裂原作用。ET-1的丝裂原作用主要由其A型受体(ETR_A)所介导,ETR_A的特异拮抗剂BQ123可显著抑制缺氧以及缺氧与ET-1协同所产生的增殖作用,而且发现ETR_A在缺氧培养的PASMC中的表达显著高于常氧对照组PASMC。本研究表明ET-1参与了缺氧性肺动脉结构重组,而缺氧可增强PASMC对ET-1的增殖反应性。 相似文献
998.
大鼠颈上神经节烟碱电流的整流及失敏 总被引:1,自引:0,他引:1
在培养的新生大鼠颈上神经节交感神经元标本上,用全细胞膜片钳技术研究了烟碱型乙酰胆碱受体(nAChR)通道的整流及失敏现象。烟碱激动剂引起的全细胞电流在膜电位为负值时随膜电位呈线性改变,而在膜电位达+60mV时仍不出现外向电流,表现出强烈的内向整流。nAChR通道电流存在失敏现象,即持续恒压喷射激动剂所引起的全细胞电流随时间呈指数衰减,不能保持在峰值水平,失敏随激剂浓度呈量效关系,膜电位的超极化也加 相似文献
999.
Aortal collagen typing in monkey and man showed the presence of types I, HI and V in human aorta and types I and III in monkey
aorta. Type III collagen was found to be a predominate type in both species. The molecular weight of type III collagen was
similar in these species while type I collagen was different. Both monkey and human collagen types I and III were found to
be immunogenic. Type I collagen was significantly increased while type III was decreased in human atherosclerotic plaque.
Collagen typing in fatty streak remained unaltered. 相似文献
1000.
The distribution of immunoreactive alpha-melanocyte stimulating hormone (alpha-MSHI) in the rat lower brain stem was examined by indirect immunofluorescence or peroxidase- anti-peroxidase immunohistochemical method using an antiserum against synthetic alpha-MSH. The results confirmed the presence of alpha-MSHI fibers in the midbrain central gray matter and parabrachial area, and demonstrated a much more extensive distribution of these fibers in various parts of the lower brain stem areas previously thought not contain alpha-MSHI fibers. In addition, the commissural nucleus was identified as a new alpha-MSHI neurons-containing site. No alpha-MSHI neurons were seen in other regions of the rat lower brain stem. 相似文献