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91.
Little is known about the cellular and molecular regulation of the uptake process of the water-soluble vitamin biotin into liver cells, the major site of biotin utilization and metabolism. Such studies are best done using a highly viable and homogeneous cellular system that allows examination of prolonged exposure to an agent(s) or a particular condition(s) on the uptake process. Isolated hepatocytes when maintained in primary culture lose their ability to transport biotin by the specialized carrier system. The aim of the present study was, therefore, to examine the mechanism(s) of biotin uptake by the cultured human-derived liver cells, Hep G2. Uptake to biotin by Hep G2 cells was appreciable and linear for up to 10 min of incubation. The uptake process was Na+ gradient-dependent as indicated by studies of Na+ replacement and pretreatment of cells with gramicidin and ouabain. Biotin uptake was also dependent on both incubation temperature and intracellular energy. Unlabeled biotin and the structural analogs with free carboxyl groups (thioctic acid, desthiobiotin) but not those with blocked carboxyl group (biocytin, biotin methyl ester, and thioctic amide) caused significant inhibition of 3H-biotin uptake at 37°C but not 4°C. Initial rate of biotin uptake was saturable as a function of concentration at 37°C but was lower and linear at 4°C. Pretreatment of Hep G2 cells with sulfhydryl group inhibitors (e.g., p-chloromer-curibenzene sulfonate) led to a significant inhibition in biotin uptake; this inhibition was effectively reversed by reducing agents (e.g., dithiothreitol). Biotin uptake was also inhibited by the membrane transport inhibitors probenecid (noncompetitively), DIDS and furosemide but not by amiloride. Pretreatment of Hep G2 cells with valinomycin did not alter biotin uptake. The stoichiometric ratio of biotin to Na+ uptake in Hep G2 cells was also determined and found to be 1:1. These findings demonstrate that biotin uptake by these cultured liver cells is mediated through a specialized carrier system that is dependent on Na+-gradient, temperature, and energy and transports the vitamin by an electroneutral process. These findings are similar to those seen with native liver tissue preparations and demonstrate the suitability of Hep G2 cells for in-depth investigations of the cellular and molecular regulation of biotin uptake by the liver. © 1994 Wiley-Liss, Inc. 1 This article is a US Government work, and as such, is in the public domain in the United State of America . 相似文献
92.
旋毛虫肌幼虫ES抗原的基因克隆及高效表达 总被引:7,自引:0,他引:7
作者对编码旋毛虫肌幼虫ES抗原的部分结构基因进行了克隆、鉴定和表达。用RNA PCR技术直接从旋毛虫肌幼虫总RNA中反转录并扩增出0.7kh的靶DNA,酶切分析后将其克隆到融合表达载体pEx3lC中。SDS—PAGE电泳表明,含重组子的大肠杆菌能够表达出一分子量为37kDa的融合蛋白(P37),后者占菌体总蛋白的22%以上,并以包含体形式存在于菌体中。经对纯化后表达蛋白的ELlSA检测,证明它能被猪旋毛虫病阳性血清和抗旋毛虫单克隆抗体识别。研究结果揭示,重组蛋白P37对于研制旋毛虫病诊断抗原和免疫抗原具有潜在的应用价值。 相似文献
93.
双价外壳蛋白基因植物表达载体的构建及马铃薯转基因植株的鉴定 总被引:10,自引:0,他引:10
在克隆了马铃薯X病毒(PVX)、马铃薯Y 病毒(PVY)和马铃薯卷叶病毒(PLRV)的外壳蛋白基因的基础上,构建同时包含PVX和PVY 与PVY 和PLRV 两个外壳蛋白基因植物表达框架的表达载体,通过农杆菌(Agrobacterium tumefaciens)介导转化烟草(Nicotianatabacum )和生产上常用的几个马铃薯(Solanum tuberosum )优良品种:“Favorita”、“虎头”、“克4”。经PCR检测证明外源基因已整合到植物的染色体上,得到批量转基因植株。在转PVX+PVY 外壳蛋白基因的烟草上接种PVX (5 μg/m L)、PVY(20 μg/m L)病毒,得到有一定抗性的植株 相似文献
94.
High percentages of micro-calli and micro-derived embryos were produced from isolated asparagus microspores at late uninucleate stage on MS liquid medium supplemented with 1.0 mg l–1 2,4-D and 0.5 mg l–1 BA. Two types of calli, namely compact callus (CC) and loose callus (LC), were found. Plantlets were regenerated via organogenesis, when these calli were transferred onto MS solid medium supplemented with 1.0 mg l–1 BA and 0.2 mg l–1 IBA 6 weeks. Embryos were produced from liquid cultured microspores, or from solid cultured micro-calli. The frequencies of haploid plant production from organogenesis and embryogenesis were compared. Effects of plant growth regulators on callus production, plantlet regeneration, and haploid plant production were tested. The combination of BA 1.0 mg l–1 and IBA 0.2 mg l–1 resulted the highest precentage of haploid plant production (7.7% from CC, 4.3% from LC).Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- IBA
3-indolybutyric acid
- BA
6-binzyladinine
- NAA
naphtalene acetic acid
- MS
Murashige and Skoog 相似文献
95.
细胞间粘附分子1的研究进展 总被引:11,自引:0,他引:11
细胞间粘附分子1(ICAM-1),又名CD54,是一种重要的细胞表面粘附分子,属免疫球蛋白超家族.它可与鼻病毒以及整合素家族成员结合,参与炎症,普通感冒,变态反应及移植排斥反应.文章就其细胞分布、表达调节、结构功能、基因工程以及临床应用进行了综述. 相似文献
96.
97.
麦秸覆盖夏玉米对其苗期生长发育的生化他感作用研究初报 总被引:8,自引:0,他引:8
麦秸覆盖夏玉米对其苗期生长发育的生化他感作用研究初报张玉铭,马永清(中国科学院石家在农业现代化研究所,050021)AllelopathicEffectofWheatStrawMulchingonSeedlingGrowthandDevelopmen... 相似文献
98.
The interfering effects of copper, zinc, and cobalt on the uptake of mugineic acid-ferric complex were studied in barley ( Hordeum vulgare , cv. Minorimugi) grown in nutrient solution. Short-term uptake experiments of 3 h were performed utilizing both ionic and mugineic acid-complex forms of each metal at two different concentrations. Copper was most effective in decreasing iron uptake when added in an ionic form at either concentration. The inhibition order at higher concentrations followed Cu(II) > Zn(II) ≥ Co(II), Co(III), which is consistent with the stability constants of these metal complexes with mugineic acid. The displacement of iron from its mugineic acid complex by these metals is suggested as a probable explanation for the decreased iron uptake. The inhibitory effect of metal complexes with mugineic acid on iron uptake was only found in cases with higher concentrations of Cu(II) and Zn(II) complexes. Deformation of the specific iron transport system in the plasma membrane due to their adsorption may be responsible for this effect. 相似文献
99.
以网织红细胞与K_(582)细胞融合形成的胞质体杂种细胞K-RRneo为研究对象,对Vimentin、Lamin蛋白与K-RRneo细胞分化排核的关系进行了研究。结果表明:随着K-RRneo细胞的分化,Vimentin mRNA、Lamin蛋白表达明显降低,与我们整装电镜观察及Western印迹分析所得到的结论相一致,证实了薛社普提出的Vimentin、Lamin蛋白在红系细胞分化排核中所起的作用,为最终阐明红系细胞分化排核机制提供了新线索。 相似文献
100.
中国稻蝗属细胞分类学研究(英文) 总被引:4,自引:0,他引:4
本文对分布于中国境内11省区,27个采集地点的八种稻蝗进行了染色体C带核型比较研究。结果表明:中华稻蝗Oxya chinensis,上海稻蝗O.shanghaiensis,无齿稻蝗O.adentata C带分布型式相似,但染色体分组型式、交叉定位数据以及异染色质含量具有差异;山稻蝗O.agauisa具有本身独特的带纹结构,与本属其它种类迥异;双带稻蝗O.bicingula染色体带型结构相似于山稻蝗及中华稻蝗类群,表明三者在进化历程中有一定联系。小稻蝗O.hyla intricata中一个亲缘种团(Sibling species group),体现在种内的形态差异及染色体带型结构的不同。显示该种近期内分化速率较高。据此,本文讨论了稻蝗属内八个种相互间的亲缘关系和分类地位,本文认为稻蝗属内务物种进化速率是不均衡的,这种状态应归因于各物种的遗传背景和环境因素的影响。 相似文献