伪狂犬病病毒闽A株gE基因去信号肽片段在Pichia pastoris中的表达

伪狂犬病病毒囊膜糖蛋白E是一种在伪狂犬病根除计划中具有重要作用的糖蛋白.将伪狂犬病病毒闽A株gE基因去信号肽片段克隆到巴斯德毕赤酵母（Pichia pastoris）表达载体pPICZαA中,获得的重组表达载体pPICZαA-FL电击转化野生型酵母菌SMD1168后,得到多株酵母工程菌SMD1168/pPICZαA-FL.经高浓度Zeocin^TM筛选、表型鉴定、工程菌的诱导表达及表达产物的鉴定,最后得到高效表达gE基因去信号肽片段的酵母工程菌SMD1168/pPICZαA-FL-7.工程菌72 h培养上清的SDS-聚丙烯酰胺凝胶电泳与蛋白质印迹结果显示,gE基因去信号肽片段表达产物大小约为80 ku,比预期的63.8 ku大.凝胶薄层扫描结合Bradford蛋白质总含量测定结果表明,表达产物占工程菌培养上清总蛋白的13.49%,表达量可达11.7 mg/L.间接ELISA结果表明重组表达产物具有良好的抗原性,能够有效地区分伪狂犬病病毒gE标准阳性与阴性血清.     

高赖氨酸蛋白基因在大肠埃希菌中的表达

从四棱豆中克隆高赖氨酸蛋白基因wblys,通过PCR扩增wblys片段,转入原核表达载体pGEX-4T-1,构建pGEX-4T-1/wblys大肠埃希菌工程菌,表达重组蛋白,IPTG诱导后,发现细菌全蛋白在44 ku(含GST标签)处多出1条明显条带。HPLC检测赖氨酸含量。诱导后菌体总赖氨酸含量比正常菌体提高15.84 mg/g。在大肠埃希菌中高效表达植物源高赖氨酸蛋白基因,为该基因在益生菌中表达提供研究工作基础。     

北方寒冷干旱地区内陆湖泊生态安全评价指标体系研究——以呼伦湖为例

呼伦湖作为我国北方寒冷干旱地区内陆湖泊的典型代表,在涵养水源、调节气候、防风固沙及维系呼伦贝尔草原生态系统平衡乃至我国北方生态安全屏障等方面发挥着不可替代的作用。准确识别呼伦湖流域主要生态安全问题、完善其生态安全评价指标体系对于科学研判呼伦湖生态环境状况、维持区域可持续发展及精准实施底线管控政策具有重要意义。在遥感影像、地面调查、实验模拟等多尺度数据分析基础上,结合历史资料收集、专家咨询、部门访谈等方式分析识别了呼伦湖区域生态安全问题及成因：（1）气候暖干化及超载放牧等因素使得草地退化显著,土地沙化、盐碱化面积扩大;（2）草原退化增加了干草入湖、大气干湿沉降及水土流失等水环境污染负荷;（3）气候暖干化造成湿地萎缩,加之长期的过度捕捞等人类活动使得渔业资源枯竭、鱼类小型化特征显著,水生生态系统稳定性下降,蓝藻水华频发。在全面诊断分析区域生态安全问题的基础上,以山水林田湖草系统方法论为理论指导,以改善呼伦湖流域生态环境质量为核心目标,依据评价指标的针对性、科学性、可操作性及系统性原则,采用压力-状态-响应（Pressure-State-Response,PSR）模型,以气象条件、人口、水资源、水环境压力及人类活动等5个一级指标构成的压力子系统,以水环境状况、水生态状况、陆域生态状况、生态系统服务功能和敏感性、景观格局及生态风险等6个指标构成的状态子系统,以生态治理措施及生态治理投入等2个指标构成的响应子系统,共同组成了一套包含13个一级指标38个二级指标的天空地一体、陆湖统筹的生态安全评价指标体系,以期为呼伦湖及北方寒冷干旱区类似湖泊的生态安全评价研究提供借鉴。     

青藏高原典型冰川小流域土壤微生物生物量及胞外酶活性分布特征

土壤微生物释放的胞外酶是决定碳(C)、氮(N)、磷(P)生物地球化学循环的关键因素,为了阐明青藏高原典型小流域土壤微生物生物量和胞外酶活性沿海拔和土层的分布特征并揭示影响该分布格局的主要养分限制状况,于2021年8月采集了青藏高原廓琼岗日冰川小流域5个海拔梯度(4900 m; 5000 m; 5100 m; 5200 m; 5300 m)中4个土壤发生层(A层：腐殖质层、E层：淋溶层、B层：淀积层和C层：母质层)的土壤样品,定量分析了土壤基本理化性质、微生物生物量、胞外酶活性等指标。结果表明：1)微生物生物量碳氮磷的海拔差异变化规律不同,随着土层加深微生物生物量碳氮磷随海拔变化越小。同时,各海拔之间微生物生物量均有随土层加深而显著降低的趋势(P<0.05);2)四种酶活性的海拔间变化规律各异,但整体呈现随海拔升高而升高的趋势且在表层(A和E层)增长趋势更明显,而且随土层加深显著降低(P<0.05);3)该区域土壤微生物受到碳和磷共同限制,土层越深限制越高,而且海拔越高C限制越强,但P限制降低;4)青藏高原典型冰川小流域土壤微生物生物量和胞外酶活性在海拔和土层之间存在较明显的...     

青海湖岸带土壤与沉积物的地化特征与细菌群落对水位上升的响应

【目的】探究青海湖岸带土壤与沉积物的地化特征与细菌群落对水位扩张的响应。【方法】从岸上至岸下沿垂直青海湖岸带方向,采集距离湖面不同高度土壤(土壤：S1、S2)、岸边不同水深表层沉积物(过渡区：E0、E6、E17)及湖心表层沉积物(沉积物：D1、D2)样品,土壤与沉积物水深(土壤水深表示为负数)从小到大的变化表征岸边土壤被淹水转变为沉积物的过程。采用地球化学分析和16SrRNA基因高通量测序技术,探究岸带土壤与沉积物样品中的地化特征与微生物群落构成。【结果】青海湖水位上升导致的生境转变对岸带土壤与沉积物的理化性质、营养水平、有机碳类型等地化特征产生显著影响。具体表现为,随着水位升高,岸带土壤与沉积物的pH、矿物结合态有机碳含量显著升高,而碳氮比值、可溶性有机碳(dissolved organic carbon,DOC)、颗粒态有机碳含量显著下降。随着水位上升,青海湖岸带被淹没土壤的细菌群落多样性下降,且群落结构发生明显变化。这种变化与环境因子变化密切相关,具体表现为,细菌群落物种丰富度指数和香农多样性指数随着水位上升呈下降趋势;活性金属结合态有机碳含量与细菌群落多样性的变化密切相关;理化...     

酵母PHO2蛋白及其变异体与PHO5USA体外的相互作用

酵母ＰＨＯ２蛋白及其变异体与ＰＨＯ５ＵＳＡ体外的相互作用杨军,敖世洲（中国科学院上海生物化学研究所分子生物学国家重点实验室,２０００３１）关键词酵母;ＰＨＯ２;突变;ＤＮＡ结合ＰＨＯ２是酵母阻遏型酸性磷酸酯酶基因转录的正调控因子［１］,由５５９个氨基...     

Models for predicting soil zinc availability for barley

Regression models are proposed for the prediction of soil zinc availability within different pH ranges. The relationship between zinc concentration in barley plants, zinc content in soils and soil pH was described using a two-dimensional diagram. Results show that the soil zinc availability is significantly affected by pH changes at pH above 6.5. Below this point, the pH effect becomes gradually less important. In the low pH range, the soil zinc availability is mainly dependent on soil zinc quantity.     

Effects of different states of sheep fetal fibroblasts as donor cells on the early development in vitro of reconstructed sheep embryos

To investigate the effects of different states of donor cells on the development of reconstructed sheep embryos, we designed five treatments of donor cells, including cell passage, cell size, serum starvation,colchicine treatment and gene transfection. Results are as follows: ( Ⅰ ) Compared with 16-18 passage cells, the morula/blastocyst rate of 5-7 passage cells as donor nuclei was significantly higher (17.3%vs. 4.9%, P＜0.05), suggesting the advantage of short-time cultured cells in supporting the development of reconstructed embryos. (Ⅱ) The morula/blastocyst rate of reconstructed embryos derived from medium cells (15-25 μm) as donor nuclei was higher than that from large cells (25-33 μm) and small cells (8-15 μm)( 20.0% vs. 8.0%, 9.7%), indicating that reconstructed embryos from medium cells had a greater potentiality to develop into morula/blastocysts than those from small or large ones. (Ⅲ) The morula/blastocyst rate of reconstructed embryos from donor cells of SS (serum starvation) was lower than that from donor cells of NSS (non-serum starvation), but no significant difference was detected between SS and NSS( 11.8% vs. 18.6%, P＞0.05). (Ⅳ) Fetal fibroblasts treated with 0.05 μmol/L colchicine exhibited a higher morula/blastocyst rate of reconstructed embryos than those treated with 0.10 μmol/L colchicine and untreated ones (27.5% vs. 12.1%, 17.1%), however, no significant difference among the three treatments was detected (P＞0.05). (Ⅴ) The morula/blastocyst rate of reconstructed embryos from fetal fibroblasts transfected with GFP gene only was 3.1%, significantly lower than that from non-transgenic cells (3.1% vs. 20.4%, P＜0.05). In conclusion, our results demonstrated that fetal fibroblasts of fewer passages, medium size could ensure a higher morula/blastocyst rate of reconstructed embryos. Serum starvation of donor cells might be unnecessary to the development of reconstructed embryos. Donor cells treated with 0.05 μmol/L colchicine could facilitate the development of reconstructed embryos. Additionally, as cells transfected with GFP gene were used as donor nuclei, adverse effect on the development of reconstructed embryos was observed. Therefore, the developmental efficiency of reconstructed embryos could be improved if proper treatments to donor cells were used.     

ERP Correlates of Proactive and Reactive Cognitive Control in Treatment-Na?ve Adult ADHD

This study investigated whether treatment naïve adults with Attention Deficit Hyperactivity Disorder (ADHD; n = 33; 19 female) differed from healthy controls (n = 31; 17 female) in behavioral performance, event-related potential (ERP) indices of preparatory attention (CueP3 and late CNV), and reactive response control (Go P3, NoGo N2, and NoGo P3) derived from a visual cued Go/NoGo task. On several critical measures, Cue P3, late CNV, and NoGo N2, there were no significant differences between the groups. This indicated normal preparatory processes and conflict monitoring in ADHD patients. However, the patients had attenuated Go P3 and NoGoP3 amplitudes relative to controls, suggesting reduced allocation of attentional resources to processes involved in response control. The patients also had a higher rate of Go signal omission errors, but no other performance decrements compared with controls. Reduced Go P3 and NoGo P3 amplitudes were associated with poorer task performance, particularly in the ADHD group. Notably, the ERPs were not associated with self-reported mood or anxiety. The results provide electrophysiological evidence for reduced effortful engagement of attentional resources to both Go and NoGo signals when reactive response control is needed. The absence of group differences in ERP components indexing proactive control points to impairments in specific aspects of cognitive processes in an untreated adult ADHD cohort. The associations between ERPs and task performance provided additional support for the altered electrophysiological responses.     

The synthesis and characterization of mixed ligand Ru(II) complexes with dipyrido(2,3-a:3′,2′-j)phenazine (dpop′) and 2,3,5,6-tetra(2-pyridyl)-pyrazine (tppz)

The synthesis of the mixed ligand mono metallic [Ru(dpop′)(tppz)]²⁺ and bimetallic [(dpop′)Ru(tppz)Ru(dpop′)]⁴⁺ (dpop′ = dipyrido(2,3-a:3′,2′-j)phenazine; tppz = 2,3,5,6 tetra-(2-pyridyl)pyrazine) complexes is described. The [Ru(dpop′)(tppz)]²⁺ complex display an intense absorption at 518 nm which is assigned to a Ru(dπ) → dpop′ (π∗) MLCT transition, and at 447 nm which is assigned to a Ru(dπ) → tppz(π∗) MLCT transition. It undergoes emission at RT in CH₃CN with λ_em = 722 nm. The bimetallic [(dpop′)Ru(tppz)Ru(dpop′)]⁴⁺ complex shows a low energy absorption shoulder near 635 nm assigned to a Ru(dπ) → tppz(π∗) MLCT transition and an intense peak at 542 nm due to Ru(dπ) → dpop′ (π∗) MLCT transition. The bimetallic complex also emits at RT in CH₃CN with λ_em = 785 nm. Cyclic voltammetry shows reversible Ru^+2/+3 oxidations at 1.68 V for the monometallic complex and Ru^+2/+3 oxidation couples at +1.94 and +1.70 V for the bimetallic complex.