苜蓿中金属肽酶对青贮过程中蛋白降解的作用

为探讨金属肽酶在苜蓿青贮过程中对蛋白的降解作用,以苜蓿（Medicago stativa L.）绿汁发酵液模拟青贮发酵过程,添加金属肽酶抑制剂后,测定发酵后第0.5、1、1.5、2、2.5、3、4、5、7和14天时绿汁发酵液的蛋白组分。结果表明,在苜蓿发酵液发酵过程中添加金属肽酶抑制剂能够降低pH值、非蛋白氮、游离氨基酸和氨态氮含量,增加乳酸含量;发酵第14天时,抑制剂处理组非蛋白氮、游离氨基酸和氨态氮质量分数为483.98 g/kg TN,256.98 g/kg TN和26.11 g/kg TN,与对照组相比分别降低28.92%,38.59%和67.08%。金属肽酶对苜蓿发酵液蛋白降解起着一定的作用;添加金属肽酶抑制剂能够提高发酵液发酵品质。     

植物中铵转运蛋白的研究进展

铵转运蛋白在众多生物中被克隆与鉴定，它是一种广泛存在于微生物、植物细胞及动物的细胞膜上主动转运铵离子的载体，分子量约为48kD，含有10-11个跨膜域。本文阐述了植物铵转运蛋白分离鉴定的过程，对于铵转运蛋白的结构、功能、基因表达调控等方面作了较详细叙述。不同氮素条件下，铵转运蛋白基因通过转录调控表现了对铵离子吸收转运的不同特点，使植物根系在较宽的浓度范围中吸收铵离子，为细胞内铵离子库的内稳态提供了理论依据。铵转运蛋白有助于作物更有效的吸收氮素，为农业生产粮食增收提供了有利保障。     

水稻生育后期根系响应氮肥调控的蛋白作用机制分析

以大穗型水稻‘金恢809’为试验材料,设置前氮后移(施氮量的30%作基肥,30%作蘖肥,30%作穗肥,10%作粒肥)及传统氮肥(施氮量的40%作基肥,30%作蘖肥,30%作穗肥)施用两种处理,并采用双向电泳技术分析了水稻生育后期根系对前氮后移调控的蛋白响应机制。通过构建的不同氮肥处理下水稻开花后15 d根系蛋白2-DE电泳图谱,并结合MALDI-TOF/MS分析,共成功鉴定到57个出现差异表达的蛋白,其中40个在前氮后移处理下上调表达,17个下调表达。依据其蛋白功能可以归为12类,主要涉及信号转导,氨同化,胁迫与防御,糖酵解,三羧酸循环,氨基酸代谢、蛋白合成及折叠等代谢途径。研究发现水稻生育后期根系GTP结合的核蛋白、GDP解离抑制因子、液泡型H+-ATP酶亚基C会响应氮肥的调控而出现表达变化,并将信号传递到电压依赖性离子通道蛋白、液泡型ATP酶亚基C以及膜联蛋白p35,进而调节根系细胞间物质运输与信息的传递。本研究结果还表明,前氮后移的施肥方式,通过适当增加水稻生育后期氮肥的施用,调节根系糖类转换、糖酵解及三羧酸循环相关蛋白的表达,增加了根系ATP的供应;通过增强根系中清除过氧化氢系统及防御相关蛋白的表达,延缓了根系的衰老;通过上调谷氨酸脱氢酶、天冬氨酸转氨酶和谷氨酞胺合成酶的表达丰度,促进了根系对土壤中氮元素的吸收与转运。研究结果对于进一步明确水稻根系响应氮素调控的分子生态特性具有重要的理论与实际意义。     

一种从薏仁种子中提取出来的抗真菌蛋白

利用离子交换色谱柱、亲和层析色谱柱、凝胶过滤柱和反相高效液相色谱柱,从薏仁（Coix lacrymajobi）种子中分离纯化出一种抗植物病原真菌的蛋白,经HPLC鉴定纯度为98.23%。经SDS-PAGE分析,该蛋白的相对分子质量为25kD。它对链格孢菌（Alternaria alternate）、绿色木霉（Trichoderma viride）和小麦赤霉（Fusarium graminearum）有明显抑制作用,其半抑制浓度分别为9.15、4.50和12.21μmol/L。观察该抗真菌蛋白对霉菌菌丝形态的影响,在透射电镜下可以看到霉菌细胞壁变厚以及变形,细胞壁和细胞膜间间隙变大,细胞壁出现孔洞,细胞内容物渗出,细胞器受到严重破坏。     

岩藻黄质通过AKT/mTOR途径调节糖酵解代谢促进人急性淋巴细胞白血病细胞凋亡的作用研究

为阐明岩藻黄质促进人急性淋巴细胞白血病(CEM/C1)细胞凋亡的作用机制,本研究采用MTT法检测岩藻黄质处理后的CEM/C1细胞活力;利用流式细胞仪测定CEM/C1细胞的早期与晚期凋亡率以及细胞周期的分布;采用试剂盒检测岩藻黄质对CEM/C1细胞中丙酮酸激酶(PK)和己糖激酶(HK)活性以及葡萄糖摄取量、三磷酸腺苷(ATP)生成量、乳酸生成量的影响;通过蛋白免疫印迹法(WB)测定CEM/C1细胞相关蛋白表达水平,并通过分子对接进行验证。结果表明,岩藻黄质对CEM/C1细胞活力具有显著的抑制效果,并呈剂量和时间依赖性;随着岩藻黄质浓度的增加,CEM/C1细胞的早期凋亡率和晚期凋亡率明显增加,G1/G2细胞的比例显著或极显著下降,S期细胞的比例显著或极显著上升;岩藻黄质显著抑制糖酵解相关酶(PK、HK)的活性以及蛋白表达;岩藻黄质显著降低磷酸化mTOR和磷酸化AKT蛋白的表达水平;岩藻黄质与AKT和mTOR蛋白对接的结合能均小于-5 kcal·mol^-1,主要通过氢键作用于酶活中心。综上,岩藻黄质能显著促进CEM/C1细胞凋亡,其作用机制可能与AKT/mTOR信号通过...     

猪血浆中多种功能蛋白的连续提取工艺

为了充分利用中国丰富的猪血资源,提高猪血产品经济效益和社会效益,该文以提取过程各阶段功能蛋白质含量变化为参考指标,对猪血浆中主要功能蛋白,如凝血酶、纤维蛋白原、白蛋白及免疫球蛋白等进行了连续提取,最终确立了一套切实可行的连续提取工艺：将柠檬酸钠抗凝的血浆经冻融处理,离心得到沉淀纤维蛋白原,在上清液中加入氯化钡,利用吸附法提取凝血酶后再经盐析法分离提取白蛋白及免疫球蛋白。1 L血浆可同时提得纤维蛋白原2.67 g、凝血酶粗品0.22 g(比活力为38.5 U/mg)、白蛋白31.41 g和免疫球蛋白13.8     

超临界CO_2杀灭大肠杆菌过程中菌体蛋白的变化

通过SDS-聚丙烯酰胺凝胶电泳,结合超临界CO2对大肠杆菌的杀菌曲线,研究超临界压力、温度和处理时间对大肠杆菌菌体蛋白的影响.结果表明:超临界CO2在杀灭大肠杆菌过程中,可显著降低菌体碱溶性(pH8.0)蛋白的溶解性,但对菌体总蛋白含量和种类没有影响;37℃下超临界CO2处理大肠杆菌30min,压力为10MPa时,大肠杆菌存活率为2.8%,菌体碱溶性蛋白的溶解性显著降低,当压力增加到50MPa时,大肠杆菌存活率和碱溶性蛋白溶解性变化不明显;超临界温度和处理时间对菌体碱溶性蛋白溶解性的影响与大肠杆菌在超临界CO2中的存活趋势基本一致,随处理时间的延长和温度的增加大肠杆菌存活率逐渐降低到0,碱溶性蛋白溶解性逐渐降低,直至部分蛋白带消失,但菌体总蛋白含量和种类并没有明显变化;升高温度比增加压力更能显著地导致菌体碱溶性蛋白变性,溶解性下降.     

巴西固氮螺菌中杂合双组分蛋白Org35与NifA的相互作用

巴西固氮螺菌(Azospirillum brasilense)可以与玉米(Zea mays)、小麦(Triticum aestivum)等重要作物联合固氮,是重要的潜在生物肥料.NifA蛋白是所有固氮基因转录所必需的激活蛋白.本研究利用融合蛋白下拉分析,探讨了Org35蛋白与NifA蛋白的相互作用.研究表明二者存在互作.Org35蛋白由3个独立结构区域组成,即PAS结构域、组氨酸蛋白激酶结构域和应答调节蛋白区域.本研究采用融合蛋白下拉分析了Org35蛋白三个结构域与NifA的互作关系,结果表明Org35蛋白与NifA蛋白之间的相互作用是由PAS结构域介导的.     

乙肝病毒X蛋白结合蛋白最新研究进展

乙肝病毒X蛋白结合蛋白（HBXIP）是肝细胞癌变过程的一个关键因素,它能在动物肌肉组织和恶性肿瘤组织中过表达。近年来研究显示：HBXIP能与人体内多种蛋白结合。本文综述了HBXIP蛋白参与细胞凋亡与增殖、细胞周期进程、中心体复制、肿瘤细胞迁移等过程,以期为以HBXIP蛋白为靶标的新的抗乙肝病毒等药物设计提供基础。     

扩展蛋白与果实衰老的关系

扩展蛋白（expansins）是近年来发现的植物细胞壁中特有的一类蛋白,它们的基础作用是缓释细胞壁结构网中的张力,使细胞壁变得松弛,在果实的生长发育中,起着极为重要的作用。叙述了扩展蛋白的特性,基因家族,作用机制,生理功能,其中重点论述了扩展蛋白与果实软化絮败的关系。     

A Population-Size Model for Protein Spot Detection in Proteomic Studies

In proteomic studies, a population of proteins are often examined on a gel using a technique called two-dimensional gel eletrophoresis. The technique separates the protein population into individual protein spots on a two-dimensional gel by isoelectric charge and molecular weight. The resulting gel images are then processed by a software system for spot detection and subsequent analysis. The performance of a spot-detection program is evaluated by the total number of spots that are detected. A popular spot-detection program uses the “master–slave” approach, where all spots on “slave images” are subsets of the spots on the “master image.” We argue that this approach potentially misses a large proportion of proteins and propose a model that quantifies the lack of performance. We provide nonparametric estimators for the protein population size and the expected number of proteins to be detected if a “fusion-gel” approach was used. Using the data from a rat liver proteome study, we estimate that more than half of the protein population is missed by the master–slave approach.     

叶蛋白专用打浆机的研究

从提取叶蛋白专用打浆机的特殊要求出发，提出叶蛋白专用打浆机的工作原理，研制了９ＤＹ－５０型叶蛋白专用打浆机，通过试验分析得出了生产率与功耗的计算公式     

A Rapid Protein Digestibility Assay for Identifying Highly Digestible Sorghum Lines

Protein digestibility in sorghum (Sorghum bicolor (L.) Moench) lines was determined using two standard procedures (pepsin digestibility and pH‐stat) and compared with a newly developed, rapid electrophoresis‐based screening assay. The new assay was based on the rate of α‐kafirin disappearance after pepsin digestion. α‐Kafirin, the major sorghum storage protein, makes up ≈60–70% of the total protein in the grain. In the new assay, samples were first digested with pepsin for 1 hr, and undigested proteins were then analyzed by SDS‐PAGE. The intensitizes of the undigested α‐kafirin bands were measured. Higher band intensity indicated lower protein digestibility. The new assay was significantly correlated with the standard pepsin digestibility assay (r = −0.96, n = 16) after which it was patterned. The same was true of the pH‐stat procedure (r = −0.85, n = 16). This implies that the new assay is comparable to existing procedures and can be used for screening sorghum lines for protein digestibility. Two groups consisting of high‐protein digestibility and wild‐type sorghum lines were identified when the new assay was tested on 48 sorghum lines derived from crosses of wild‐type and mutant high protein digestibility lines, indicating that the new assay was efficient in differentiating between the two groups. Advantages of the new assay over the standard procedures include considerable reduction in analysis time and sample size required for the analysis. For example, analysis time was reduced by 20% and sample size by 10% when the new assay was used as compared with the pH‐stat procedure. We estimate that ≈60 sorghum lines can be screened in a day by a single operator using the new assay.     

Role for p53 in selenium-induced senescence

The tumor suppressor p53 and the ataxia-telangiectasia mutated (ATM) kinase play important roles in the senescence response to oncogene activation and DNA damage. It was previously shown that selenium-containing compounds can activate an ATM-dependent senescence response in MRC-5 normal fibroblasts. Here, the shRNA knockdown approach and other DNA damage assays are employed to test the hypothesis that p53 plays a role in selenium-induced senescence. In MRC-5 cells treated with methylseleninic acid (MSeA, 0-10 μM), depletion of p53 hampers senescence-associated expression of β-galactosidase, disrupts the otherwise S and G2/M cell cycle arrest, desensitizes such cells to MSeA treatment, and increases genome instability. Pretreatment with KU55933, an ATM kinase inhibitor, or NU7026, an inhibitor of DNA-dependent protein kinase, desensitizes MSeA cytotoxicity in scrambled but not p53 shRNA MRC-5 cells. These results suggest that p53 is critical for senescence induction in the response of MRC-5 noncancerous cells to selenium compounds.     

昆虫化学感受蛋白及其功能研究进展

化学感受蛋白(chemosensory protein,CSP)是一类广泛表达于昆虫体内的水溶性小分子量蛋白,1994年首次从黒腹果蝇(Drosophila melanogaster)中克隆报道,随后在几乎所有昆虫类群中都被广泛鉴定,CSP编码基因在物种基因组中多以基因簇形式存在,表明其源于相同祖先。本文对CSP理化特性、表达特点、结构特征和进化模式等研究进行概述,重点介绍了CSP功能研究的主要成果。根据本实验室该方向的研究,结合他人已有研究成果,推导出了昆虫CSP的新功能,即CSP具有保护和运输在代谢过程有重要作用的小分子化合物的功能。     

Role of phospholipase A2 in the induction of drip loss in porcine muscle

The role of phospholipase A2 in the induction of drip loss from pig muscle has been investigated. In samples from porcine M. longissimus dorsi, total PLA2 activity as well as mRNA and protein levels of the group VIA iPLA2 (iPLA2-VIA) increased during the initial 4 h post-mortem period. Morphological studies of porcine muscle showed that at 4 h post-mortem, gaps had formed between muscle fibers and that the sarcolemma membrane borders appeared blurred. At the same time iPLA2-VIA protein levels were increased inside muscle fibers and at the sarcolemma. iPLA2-VIA mRNA abundance in samples from different breeds of pigs with variations in drip loss revealed no clear correlation between drip loss level and iPLA2-VIA expression. Together, these data indicate that during the post-mortem period, iPLA2-VIA expression and activity is increased at the muscle fiber membranes. PLA2 activity may affect membrane permeability and consequently the progression of drip formation in porcine muscle.     

Role of root-induced changes in the rhizosphere for iron acquisition in higher plants

Plants can mobilize iron (Fe) in the rhizosphere by non-specific and specific (adaptive) mechanisms. Non-specific mechanisms are, for example, rhizosphere acidification related to high cation-anion uptake ratios, or citric acid excretion. The specific mechanisms are root responses to Fe deficiency and can be classified into two different strategies. The Strategy I is typical for dicots and monocots except for grasses (graminaceous species) and is characterized by increased plasma membrane-bound reductase activity, enhanced net excretion of protons and enhanced release of reducing compounds, mainly phenolics. The reductase activity is stimulated by low pH, and with supply of Fe^III chelates, ferric reduction at the plasma membrane takes place prior to uptake. In contrast, in graminaceous species (Strategy II) these root responses are absent, but enhancement of release of Fe^III chelating compounds - phytosiderophores - takes place. These phytosiderophores are very efficient in mobilizing Fe^III from artificially prepared sparingly soluble inorganic compounds (e.g. Fe^III hydroxide) and from calcareous soils. The ferrated phytosiderophores are taken up by grasses at rates 10² to 10³ times higher than Fe supplied either as synthetic chelate or microbial siderophores (e.g. ferrioxamine B), indicating a specific membrane transport system for ferrated phytosiderophores in roots of grasses. In calcareous soils phytosiderophores not only mobilize Fe, but also Zn, Mn, and Cu by chelation. However, only the Fe^III phytosiderophores are taken up preferentially by Fe deficient grasses. The ecological advantages and disadvantages of Strategy I and Strategy II for Fe acquisition from calcareous soils are discussed.     

用于药用蛋白生产的外源表达系统

利用外源蛋白表达系统生产具有重要价值的药用蛋白是现代生物技术产业的核心内容和研究热点,也是各国政府和制药企业竞相巨额资助的研究领域之一。本文综合分析了目前用于重组药用蛋白生产的原核表达系统、酵母表达系统、昆虫表达系统、哺乳动物表达系统和植物表达系统在表达产量、翻译后加工能力、生产周期、生产成本、适用范围等方面的优劣以及研究现状和最新研究进展,揭示出植物表达系统将在未来重组蛋白大规模生产中占据重要地位。但是现有的各种表达系统都无法单独实现各种重组蛋白高活性、低成本、安全系数高的大规模生产。在相当长时间里,各种表达系统在重组蛋白生产上将长期共存。而通过遗传改造、基因组学、蛋白质组学等研究方法不断改进各种外源蛋白表达系统的性能,不断建立更加优越的外源蛋白表达系统则是大家共同努力的目标。