SSR标记辅助回交转育大豆7S球蛋白α-亚基致敏蛋白缺失新品系

为快速培育7S球蛋白致敏蛋白α-亚基缺失的优质大豆新品系,以（α＇＋α）-亚基双缺失型材料日B为供体亲本,东农47为受体亲本,以杂交一回交转育方法为基础,结合SSR标记辅助遗传背景选择与主要农艺性状鉴定及品质分析,在BC2F4选育到7S球蛋白α-亚基缺失新品系Cb80。Cb80综合农艺性状优良,遗传背景回复率大于90％,其蛋白质及氨基酸总量为44．1％、41．95％,分别比轮回亲本东农47提高3．5和3．76个百分点。     

大米蛋白的酶水解机制研究——Ⅱ酶水解过程中蛋白质的组分变化

凝胶色谱和高效液相色谱分析表明,大米蛋白在碱性蛋白酶Alcalase水解过程中,其可溶性蛋白组分的相对分子质量(Mr.)分布范围相对稳定,但Mr.1350u以下小分子组分的相对含量不断增加;十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析显示,随着酶水解反应的进行,Mr.为57、39、26、22ku的亚基逐渐消失,而29ku和13ku两个亚基的含量相应增加,且这两个亚基表现出可抵抗酶水解的特性。氨基酸分析表明,酶解后残余物蛋白中胱氨酸、蛋氨酸等含硫氨基酸的含量显著高于可溶性蛋白中的含量。圆二色光谱(CD)分析显示,酶水解前后大米蛋白的二级结构发生显著变化,在残余物蛋白质组分中自由回转的比例高达71.3%,α-螺旋结构完全消失。     

调控乙酰CoA合成酶表达促进乙酸利用及GlcNAc合成

N-乙酰氨基葡萄糖(GlcNAc)是一种重要的功能单糖。在利用枯草芽孢杆菌发酵产GlcNAc的过程中,乙酸的积累严重抑制了细胞生长和GlcNAc合成。为了获得高效利用乙酸生产GlcNAc的微生物细胞工厂,作者通过突变乙酰CoA(Ac-CoA)合成酶编码基因acsA 5′UTR区域内全局转录调控因子CodY结合序列,调控acsA转录水平,进一步突变AcsA乙酰化调控位点,解除乙酰化修饰,促进乙酸转化为Ac-CoA,最终乙酸积累量由6.3 g/L减少到3.6 g/L,同时细胞干重(DCW)由2.7 g/L增加到5.3 g/L,GlcNAc产量由1.9 g/L提高到5.0 g/L,为进一步获得高产GlcNAc细胞工厂提供了基础。     

碱性蛋白酶水解对豆粕中大豆抗原蛋白的影响

大豆是八大食物过敏源之一,其中主要蛋白成分7S和11S球蛋白也是致敏性较强的抗原蛋白。本试验研究了豆粕在Alcalase酶水解过程中7S和11S两种大豆抗原蛋白的变化。结果表明,酶解10 min时7S伴球蛋白的α’、α和β亚基、酶解60 min时11S球蛋白的酸性亚基等主要抗原蛋白成分即被完全水解,同时新产生了23 ku和大量14 ku以下组分;酶解豆粕中95.1%的蛋白可溶于水,其中68.1%蛋白质可溶于三氯乙酸。     

燕麦蛋白组分分离提取及其SDS-PAGE电泳分析

通过单因素实验对燕麦蛋白组分的分离提取工艺进行了优化,并通过SDS-PAGE电泳对燕麦蛋白组分进行亚基分析。结果表明:燕麦清蛋白提取的最佳温度为40℃,球蛋白提取最佳盐浓度为7%,醇溶蛋白提取的最佳乙醇浓度为75%,谷蛋白提取的最佳碱浓度为0.05 mol/L,蛋白质提取率为83.1%。SDS-PAGE实验结果显示:燕麦清蛋白在10~100 kD范围内均有分布,燕麦球蛋白由2个亚基组成,分子量分别在97.4~100 kD和43~66.2 kD范围内,燕麦醇溶蛋白亚基大部分集中在18.39~40.72kD之间,燕麦谷蛋白部分亚基分布在20.67~26.66kD与43.29~50.80 kD之间。     

Adaptation of Oxidative Phosphorylation Machinery Compensates for Hepatic Lipotoxicity in Early Stages of MAFLD

Alterations in mitochondrial function are an important control variable in the progression of metabolic dysfunction-associated fatty liver disease (MAFLD), while also noted by increased de novo lipogenesis (DNL) and hepatic insulin resistance. We hypothesized that the organization and function of a mitochondrial electron transport chain (ETC) in this pathologic condition is a consequence of shifted substrate availability. We addressed this question using a transgenic mouse model with increased hepatic insulin resistance and DNL due to constitutively active human SREBP-1c. The abundance of ETC complex subunits and components of key metabolic pathways are regulated in the liver of these animals. Further omics approaches combined with functional assays in isolated liver mitochondria and primary hepatocytes revealed that the SREBP-1c-forced fatty liver induced a substrate limitation for oxidative phosphorylation, inducing enhanced complex II activity. The observed increased expression of mitochondrial genes may have indicated a counteraction. In conclusion, a shift of available substrates directed toward activated DNL results in increased electron flows, mainly through complex II, to compensate for the increased energy demand of the cell. The reorganization of key compounds in energy metabolism observed in the SREBP-1c animal model might explain the initial increase in mitochondrial function observed in the early stages of human MAFLD.     

55SiMnMo钢正火组织的精细结构和晶体学特征

55SiMnMo 钢的正火组织由无碳化物上、下贝氏体,马氏体和残余奥氏体组成。贝氏体含有中脊线,组织单元和位错;马氏体含有孪晶;残余奥氏体含有高密度位错。奥氏体的位向关系和惯习面表明它以切变机制生成。从界面富碳和界面应力弛豫两方面阐述了贝氏体组织单元以及贝氏体片条的生成机制。     

The Pro-Inflammatory Deletion Allele of the NF-κB1 Polymorphism Is Characterized by a Depletion of Subunit p50 in Sepsis

The functionally important NF-κB1 promoter polymorphism (−94ins/delATTG) significantly shapes inflammation and impacts the outcome of sepsis. However, exploratory studies elucidating the molecular link of this genotype-dependent pattern are lacking. Accordingly, we analyzed lipopolysaccharide-stimulated peripheral blood mononuclear cells from both healthy volunteers (n = 20) and septic patients (n = 10). All individuals were genotyped for the −94ins/delATTG NF-κB1 promoter polymorphism. We found a diminished nuclear activity of the NF-κB subunit p50 in ID/DD genotypes after 48 h of lipopolysaccharide stimulation compared to II genotypes (p = 0.025). This was associated with higher TNF-α (p = 0.005) and interleukin 6 concentrations (p = 0.014) and an increased production of mitochondrial radical oxygen species in ID/DD genotypes (p = 0.001). Although ID/DD genotypes showed enhanced activation of mitochondrial biogenesis, they still had a significantly diminished cellular ATP content (p = 0.046) and lower mtDNA copy numbers (p = 0.010) compared to II genotypes. Strikingly, these findings were mirrored in peripheral blood mononuclear cells taken from septic patients. Our results emphasize the crucial aspect of considering NF-κB subunits in sepsis. We showed here that the deletion allele of the NF-κB1 (−94ins/delATTG) polymorphism was associated with the lower nuclear activity of subunit p50, which, in turn, was associated with aggravated inflammation and mitochondrial dysfunction.     

就地γ谱仪测量某坑道内222Rn-220Rn子体剂量比

为了解某坑道内222Rn-220Rn子体α潜能浓度比及剂量比的变化,使用就地HPGeγ谱仪分别在坑道内密闭环境和通风两种状态下进行测量,使用相对效率法和α潜能浓度法对测量结果进行分析。结果表明:测量期间,密闭坑道内222Rn-220Rn子体α潜能浓度比平均为3.6,子体剂量比平均为10.8;通风8小时内,坑道中222Rn-220Rn子体α潜能浓度比平均为1.2,子体剂量比平均为3.6;通风约14小时后,坑道内222Rn-220Rn子体α潜能浓度比平均为0.2,子体剂量比平均为0.6,220Rn子体剂量占222Rn-220Rn子体总剂量份额的62%。使用就地HPGeγ谱仪可快速、连续得到坑道内222Rn-220Rn子体α潜能浓度比及剂量比,在未来的222Rn-220Rn剂量研究中有着广阔的应用前景。     

Improvement of nutritional value and functional properties of soybean glycinin by protein engineering

Glycinin is one of the predominant storage proteins of soybean.To improve its functional properties (heat-induced gelationand emulsification) and/or nutritional value, the A_1aB_1b proglycininsubunit was modified on the basis of genetically variable domainssuggested from the comparison of amino acid sequences of glycinin-typeglobulins from various legumes and nonlegumes and the relationshipsbetween the structure and the functional properties of glycinin.Thus, nucleotide sequences corresponding to each of the variabledomains were deleted from the cDN A encoding the A_1aB_1b proglycinin,and a synthetic DNA encoding four continuous methionines wasinserted into the cDNA region corresponding to each of the variabledomains. Expression plasmids carrying the modified cDNAs wereconstructed and expressed in Escherichia coli strain JM105.Some of the modified proteins were accumulated as soluble proteinsin the cells at a high level and self-assembled. They exhibitedfunctional properties superior to those of the native glycininfrom soybean, which establishes the possibility of creatingtheoretically designed novel glycinins with high food qualities.