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21.
珍珠黄杨叶片的蛋白质提取方法探讨 总被引:3,自引:0,他引:3
蛋白质样品制备是双向电泳的核心.为了找到一种适合提取珍珠黄杨叶片蛋白质的方法,本文以该树种扦插苗的叶片为材料,用TCA-丙酮沉淀法、Tris-饱和酚法和2-D Clean-up Kit提取蛋白质,并进行双向凝胶电泳,采用银染法进行检测.结果表明,TCA-丙酮沉淀法得到的样品图谱背景模糊、拖尾;Tris-饱和酚法得到的样品图谱清晰,蛋白点饱满,无纵向或横向拖尾,但有蛋白点丢失;2-D Clean-Up Kit提取的蛋白质样品得到了较好双向电泳图谱. 相似文献
22.
Manman Zhang Wenliang Gong Dianjun Zhang Ming Ji Binjie Chen Beina Chen Xinyu Li Yuefei Zhou Chengyi Dong Gehua Wen Xiaoni Zhan Xiafang Wu Lulu Cui Yuliang Feng Siman Wang Huiya Yuan Enyu Xu Maosheng Xia Alexei Verkhratsky Baoman Li 《Cell death & disease》2022,13(4)
Alzheimer’s disease (AD) is the prevalent cause of dementia in the ageing world population. Apolipoprotein E4 (ApoE4) allele is the key genetic risk factor for AD, although the mechanisms linking ApoE4 with neurocognitive impairments and aberrant metabolism remains to be fully characterised. We discovered a significant increase in the ApoE4 content of serum exosomes in old healthy subjects and AD patients carrying ApoE4 allele as compared with healthy adults. Elevated exosomal ApoE4 demonstrated significant inverse correlation with serum level of thyroid hormones and cognitive function. We analysed effects of ApoE4-containing peripheral exosomes on neural cells and neurological outputs in aged or thyroidectomised young mice. Ageing-associated hypothyroidism as well as acute thyroidectomy augmented transport of liver-derived ApoE4 reach exosomes into the brain, where ApoE4 activated nucleotide-binding oligomerisation domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome by increasing cholesterol level in neural cells. This, in turn, affected cognition, locomotion and mood. Our study reveals pathological potential of exosomes-mediated relocation of ApoE4 from the periphery to the brain, this process can represent potential therapeutic target.Subject terms: Cognitive neuroscience, Alzheimer''s disease, Cellular neuroscience 相似文献
23.
Characterization of an Escherichia coli O157:H7 O-Antigen Deletion Mutant and Effect of the Deletion on Bacterial Persistence in the Mouse Intestine and Colonization at the Bovine Terminal Rectal Mucosa 下载免费PDF全文
Haiqing Sheng Ji Youn Lim Maryann K. Watkins Scott A. Minnich Carolyn J. Hovde 《Applied microbiology》2008,74(16):5015-5022
24.
Some properties of the β-N-acetyl-D-hexosaminidase purified from intercellular fluid of tomato leaves after the plant was systematically infected by TMV (tobacco mosaic virus) were studied. When pNP β-D-GlcNAc (p nitrophenyl-N-aeetyl β-D-glucosaminide) or pNP β-D- GalNAc (p-nitrophenyl-N-acetyl-β-D galactosaminide) was used as the substrate, it showed the optical pH between 4. 8--5.0 and optical temperature between 44— 47℃. Studies of thermostabillty indicated that the enzyme had a biphasic denaturation curve. Using pNP-β-D-GIcNAc or pNP-β-D GalNAc as the substrate, the Km value of the enzyme was 0. 36 and 0. 67 mmol/L respectively. N acetyi-D glucosamine and N acetyl-D-galactosamine were competitive inhibitors of the enzyme activities. Ag+ and Hg2+ were sensitive inhibitors and Fe2+ . Fe3+ and Cu2+ were also inhibitors enzyme activities. 相似文献
25.
Martina Kyselková Jiří Janata Markéta Ságová-Marečková Jan Kopecký 《Archives of microbiology》2010,192(3):195-200
In acetohydroxy acid synthase from Streptomyces cinnamonensis mutants affected in valine regulation, the impact of mutations on interactions between the catalytic and the regulatory subunits
was examined using yeast two-hybrid system. Mutations in the catalytic and the regulatory subunits were projected into homology
models of the respective proteins. Two changes in the catalytic subunit, E139A (α domain) and ΔQ217 (β domain), both located
on the surface of the catalytic subunit dimer, lowered the interaction with the regulatory subunit. Three consecutive changes
in the N-terminal part of the regulatory subunit were examined. Changes G16D and V17D in a loop and adjacent α-helix of ACT
domain affected the interaction considerably, indicating that this region might be in contact with the catalytic subunit during
allosteric regulation. In contrast, the adjacent mutation L18F did not influence the interaction at all. Thus, L18 might participate
in valine binding or conformational change transfer within the regulatory subunits. Shortening of the regulatory subunit to
107 residues reduced the interaction essentially, suggesting that the C-terminal part of the regulatory subunit is also important
for the catalytic subunit binding. 相似文献
26.
Hye Min Kim Min Jin Lee Ji Young Jung Chung Yeon Hwang Mincheol Kim Hee-Myong Ro Jongsik Chun Yoo Kyung Lee 《Journal of microbiology (Seoul, Korea)》2016,54(11):713-723
The increasing temperature in Arctic tundra deepens the active layer, which is the upper layer of permafrost soil that experiences repeated thawing and freezing. The increasing of soil temperature and the deepening of active layer seem to affect soil microbial communities. Therefore, information on soil microbial communities at various soil depths is essential to understand their potential responses to climate change in the active layer soil. We investigated the community structure of soil bacteria in the active layer from moist acidic tundra in Council, Alaska. We also interpreted their relationship with some relevant soil physicochemical characteristics along soil depth with a fine scale (5 cm depth interval). The bacterial community structure was found to change along soil depth. The relative abundances of Acidobacteria, Gammaproteobacteria, Planctomycetes, and candidate phylum WPS-2 rapidly decreased with soil depth, while those of Bacteroidetes, Chloroflexi, Gemmatimonadetes, and candidate AD3 rapidly increased. A structural shift was also found in the soil bacterial communities around 20 cm depth, where two organic (upper Oi and lower Oa) horizons are subdivided. The quality and the decomposition degree of organic matter might have influenced the bacterial community structure. Besides the organic matter quality, the vertical distribution of bacterial communities was also found to be related to soil pH and total phosphorus content. This study showed the vertical change of bacterial community in the active layer with a fine scale resolution and the possible influence of the quality of soil organic matter on shaping bacterial community structure. 相似文献
27.
28.
MATE (multidrug and toxic compound extrusion) transporter proteins mediate metabolite transport in plants and multidrug resistance in bacteria and mammals. MATE transporter NorM from Vibrio cholerae is an antiporter that is driven by Na+ gradient to extrude the substrates. To understand the molecular mechanism of Na+‐substrate exchange, molecular dynamics simulation was performed to study conformational changes of both wild‐type and mutant NorM with and without cation bindings. Our results show that NorM is able to bind two Na+ ions simultaneously, one to each of the carboxylic groups of E255 and D371 in the binding pocket. Furthermore, this di‐Na+ binding state is likely more efficient for conformational changes of NorM_VC toward the inward‐facing conformation than single‐Na+ binding state. The observation of two Na+ binding sites of NorM_VC is consistent with the previous study that two sites for ion binding (denoted as Na1/Na2 sites) are found in the transporter LeuT and BetP, another two secondary transporters. Taken together, our findings shed light on the structure rearrangements of NorM on Na+ binding and enrich our knowledge of the transport mechanism of secondary transporters. Proteins 2014; 82:240–249. © 2013 Wiley Periodicals, Inc. 相似文献
29.
大鼠实验性脾虚证胰腺组织化学研究 总被引:6,自引:0,他引:6
用成年雄性Wistar大鼠30只,分为(1)正常对照组,喂饲自来水。(2)脾虚组,用苦降破气中药和饮食失节法致成脾虚模型。(3)自然恢复组,动物致虚后,喂饲自来水。(4)中药治疗组。取四组动物胰腺进行RNA,琥珀酸脱氢酶(SDH),乳酸脱氢酶(LDH),三磷酸腺苷酶(ATPase),葡萄糖-6-磷酸酶(G-6-Pase)和硫胺素焦磷酸酶(TPPase)组织化学反应和观察,并对SDH,LDH,RNA进行了显微分光光度计定量测定。本研究结果表明,脾虚组胰腺泡细胞的RNA,SDH,ATPase,G-6-Pase,TPPase含量和活性都低于对照组,而LDH活性高于对照组。治疗组与自然恢复组相比,治疗组胰腺泡细胞以上指标接近对照组。定量测定与定性的结果一致。本研究表明,脾虚证时胰腺泡细胞上述几种酶活性和RNA明显下降,可能在脾虚证发病中起主要作用,中药治疗有显著改善 相似文献
30.
Jiří Váňa 《Plant Systematics and Evolution》1970,118(5):572-579
Summary A second species of the hitherto monotypic genusKymatocalyx
Herz. is described and figured in detail. The new combinationKymatocalyx dominicensis (Spruce) comb. nova is proposed. 相似文献