利用单链构象多态性检测牛白细胞黏附缺陷症方法的建立

牛白细胞黏附缺陷症(BLAD)是由常染色体上CD18基因单碱基突变(A→G)引起的隐性遗传疾病,隐性基因纯合时导致白细胞表面的β2整合素表达明显减少或缺乏而引起临床发病,患病牛的主要特征是机体免疫力降低、易患病从而影响其生产性能的表现,严重影响了奶牛场的经济效益。该工作利用自行设计的特异性PCR引物,通过对PCR、PCR-SSCP条件的筛选和优化,并结合DNA序列测定,建立了PCR-SSCP检测BLAD 有害基因的新方法,该方法简便快捷、准确率高,使用样品宽泛,适合大样本筛选,为奶牛BLAD有害基因的分型和筛选提供了新的方法和思路,为我国奶牛的分子选育提供了可靠的理论依据。     

谷氨酸棒杆菌碱基编辑的条件优化

近年来,基于CRISPR/Cas9的碱基编辑技术因其具有不产生DNA双链断裂、无需外源DNA模板、不依赖宿主同源重组修复的优势,已经逐渐发展成为一种强大的基因组编辑工具,在动物、植物、酵母和细菌中得到了开发和应用。研究团队前期已在重要的工业模式菌株谷氨酸棒杆菌中开发了一种多元自动化的碱基编辑技术MACBETH,为进一步优化该方法,提高碱基编辑技术在谷氨酸棒杆菌中的应用效率,本研究首先在谷氨酸棒杆菌中构建了基于绿色荧光蛋白(GFP)的检测系统:将GFP基因的起始密码子ATG人工突变为ACG,GFP无法正常表达,当该密码子的C经编辑后恢复为T,即实现GFP蛋白的复活,结合流式细胞仪分析技术,可快速衡量编辑效率。然后,构建针对靶标位点的碱基编辑工具,经测试,该位点可成功被编辑,在初始编辑条件下碱基编辑效率为(13.11±0.21)%。在此基础上,通过对不同培养基类型、诱导初始OD600、诱导时间、诱导物浓度进行优化,确定最优编辑条件是:培养基为CGXII,初始OD600为0.05,诱导时间为20 h,IPTG浓度为0.01 mmol/L。经过优化,编辑效率达到(30.35±0.75)%,较初始条件提高了1.3倍。最后,选取原编辑条件下编辑效率较低的位点,进行了优化后编辑条件下的编辑效率评估,结果显示,不同的位点在最优编辑条件下的编辑效率提高了1.7–2.5倍,进一步证实该优化条件的有效性及通用性。研究结果为碱基编辑技术在谷氨酸棒杆菌中更好的应用提供了重要的参考价值。     

Control and optimization of nitrifying communities for nitritation from domestic wastewater at room temperatures

To achieve nitritation from complete-nitrification seed sludge at room temperature of 19 ± 1 °C, a lab-scale sequencing batch reactor (SBR) treating domestic wastewater with low C/N ratios was operated to investigate the control and optimization of nitrifying communities. Ammonia oxidizing bacteria (AOB) dominance was enhanced through the combination of low DO concentrations (<1.0 mg/L) and preset short-cycle control of aeration time. Nitritation was successfully established with NO₂^?-N/NO_x^?-N over 95%. To avoid the adverse impact of low DO concentrations on AOB activities, DO concentrations were increased to 1–2 mg/L. At the normal DO levels and temperatures, on-line control strategy of aerobic durations maintained the stability of nitritation with nitrite accumulation rate over 95% and ammonia removal above 97%. Fluorescence in-situ hybridization (FISH) analysis presented that the maximal percentage of AOB in biomass reached 10.9% and nitrite oxidizing bacteria (NOB) were washed out.     

Determinants of 14-3-3σ Protein Dimerization and Function in Drug and Radiation Resistance

Many proteins exist and function as homodimers. Understanding the detailed mechanism driving the homodimerization is important and will impact future studies targeting the “undruggable” oncogenic protein dimers. In this study, we used 14-3-3σ as a model homodimeric protein and performed a systematic investigation of the potential roles of amino acid residues in the interface for homodimerization. Unlike other members of the conserved 14-3-3 protein family, 14-3-3σ prefers to form a homodimer with two subareas in the dimeric interface that has 180° symmetry. We found that both subareas of the dimeric interface are required to maintain full dimerization activity. Although the interfacial hydrophobic core residues Leu¹² and Tyr⁸⁴ play important roles in 14-3-3σ dimerization, the non-core residue Phe²⁵ appears to be more important in controlling 14-3-3σ dimerization activity. Interestingly, a similar non-core residue (Val⁸¹) is less important than Phe²⁵ in contributing to 14-3-3σ dimerization. Furthermore, dissociating dimeric 14-3-3σ into monomers by mutating the Leu¹², Phe²⁵, or Tyr⁸⁴ dimerization residue individually diminished the function of 14-3-3σ in resisting drug-induced apoptosis and in arresting cells at G₂/M phase in response to DNA-damaging treatment. Thus, dimerization appears to be required for the function of 14-3-3σ.     

华西雨屏区香樟人工林土壤表层细根生物量和碳储量

2010年11月-2011年12月, 研究了华西雨屏区31年生香樟人工林土壤表层(0～30 cm)细根生物量及碳储量.结果表明: 香樟人工林土壤0～30 cm层细根总生物量(活根+死根)和碳储量的平均值分别为1592.29 kg·hm^-2和660.68 kg C·hm^-2,其中活细根贡献率分别为91.1%和91.8%.随着土壤深度的增加,香樟1～5级活细根和死细根的生物量及碳储量均显著减少;随着根序等级的升高,香樟活细根生物量及碳储量显著增加.香樟细根总生物量及碳储量均在秋季最高、冬季最低,死细根生物量及碳储量为冬季最高、夏季最低;1级根和2级根生物量及碳储量均在夏季最高、冬季最低,而3~5级根则为秋季最高、冬季最低.土壤养分和水分的空间异质性是导致细根生物量和碳储量变化的主要原因.     

贡嘎山原始森林区苔藓植物重金属含量及其对汞的吸附特征

采集了贡嘎山海螺沟原始森林区的7种主要苔藓,分析了其重金属（Hg、Cr、Cd、Ni、Pb、Cu、Mn、Zn和Fe）含量,并选择两种分布较广的赤茎藓和多枝砂藓进行汞吸附试验.结果表明,贡嘎山海螺沟原始森林区7种苔藓的重金属含量均较低,除Cd含量与欧美地区背景值相当外,其他均低于欧美地区的背景含量,表明该地区大气未受重金属污染,大气环境质量良好. 两种苔藓对汞的吸附是一个主动快速的过程,2 h左右即达到吸附平衡,其吸附特征能很好地用Langmuir和Freundlich方程拟合,由Langmuir方程计算的两种苔藓对汞的最大吸附量分别为15.24和8.19 mg·g^-1,表明它们具有较强的汞吸附能力,可用作大气汞污染的指示生物.     

用彗星实验技术分析MTX对小鼠细胞DNA的损伤作用

MTX是一种抗叶酸药物 ,作用于增殖细胞 ,为了解其作用机制和探测其遗传毒性靶器官 ,以小鼠为研究对象 ,用彗星实验技术检测了MTX腹腔注射染毒后对脾、骨髓、胸腺、和外周血淋巴细胞的DNA损伤作用及其与MTX剂量间的相关。 1.2 5～ 5mg/kgMTX可诱发小鼠体内 4种细胞的DNA单链断裂 ,核DNA损伤程度与用药剂量呈正相关。不同种类细胞对MTX的易感性不同 ,脾、骨髓、胸腺、外周血淋巴细胞可能是MTX的遗传毒性靶细胞。外周血淋巴细胞在SCGE分析中的拖尾现象可作为用药后组织器官对药物敏感性反映的生物标志     

PAC对谷子花后土壤氮素供应和叶片抗氧化特性的影响

全基施肥方式会造成作物全生育期内营养供应失衡,导致生育后期缺氮早衰。为探究聚天门冬氨酸和壳聚糖复配剂(PAC)保障谷子(Setariaitalica)花后氮素供应和调控叶片抗氧化特性的机制,建立全基施肥背景下东北春谷防衰增产的生产技术,于2020–2021年在中国农业科学院作物科学研究所公主岭试验站开展大田试验,以谷子品种张杂谷13号和华优谷9号为材料,设置常规氮素(CN)和PAC配合氮素(PN) 6个氮素水平(0、75、112.5、150、225和337.5 kg·hm^–2)播种前进行全基施肥处理。结果表明,与常规氮肥处理相比,相同施氮量下,PAC处理后,两品种谷子花期和灌浆中期0–20cm和20–40 cm土层土壤硝态氮和铵态氮含量升高,花后叶面积显著增大,叶面积降幅减小;花后0–40天旗叶超氧化物歧化酶、过氧化物酶及过氧化氢酶活性升高,丙二醛含量降低。因此, PAC有效保障了谷子生育中、后期土壤氮素的供应,提高了叶片抗氧化能力,延缓了叶片衰老进程,进而提高产量。2020年和2021年Z13的增产幅度分别为11.24%–21.55%和8.65%–14.22%,...     

金丝猴(RHINOPITHECUS)脑的外部形态

本文是对三种金丝猴脑的外部形态的观察结果表明:金丝猴的大脑皮层除存在猴科固有的全部沟裂外,还较其他猴科动物具有更多的副沟。大脑的沟型与疣猴亚科的特征完全吻合,但是很多特点都较叶猴更近似于长臂猿。另外,除了蚓叶和蚓结节较小,四叠体的下丘较猕猴发达以外,金丝猴的小脑和脑干均与猴科的一般特征无明显差异。     

Subcellular localization of proteins of Oryza sativa L. in the model tobacco and tomato plants

The cellular localization and molecular interactions are indicative of functions of a protein. The development of a simple and efficient method for subcellular localization of a protein is indispensable to elucidate gene function in plants. In this study, we assessed the feasibility of Agrobacterium-mediated transformation (agroinfiltration) of tobacco and tomato leaf tissue to follow intracellular targeting of proteins from rice fused to green fluorescent protein (GFP). For this, a simple in planta assay for subcellular localization of rice proteins in the heterologous host systems of tobacco and tomato leaf via transient transformation was developed. We have tested the applicability of this method by expressing GFP fusions of the putative antiphagocytic protein 1 (APP1) (OsAPP, LOC_Os03g56930) and ZOS3-18-C2H2 zinc-finger protein (OsZF1, LOC_Os03g55540) from Oryza sativa L. subsp. japonica in tobacco and tomato leaf tissues. Our results demonstrate the suitability of GFP as a reporter in gene expression studies in tomato cv. MicroTom. The use of GFP-fused proteins from rice for subcellular targeting in the heterologous hosts of tobacco and tomato plant systems has been confirmed.Key words: agroinfiltration, confocal microscopy, GFP fusion protein, tomato cv, microtom