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...     

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.     

蓝藻有机无机复混肥对几种作物的增效试验

研究了几种专用蓝藻有机无机复混肥,并对农作物进行了田间肥效试验。结果表明,施用蓝藻有机无机复混肥与普通N、P、K复合肥相比较,对烟草,西芹,韭菜,康乃馨等都有增产和改良土壤的效果。烟草试验表明,蓝藻复混肥的施用量以900kg/hm2效果最好,烟草产量、产值、上等烟比例分别比对照增加7.28%,5.04%,19.81%;西芹试验表明,施用蓝藻有机无机复混肥的各处理比对照高,其中以900kg/hm2和1200kg/hm2最高;其次是600kg/hm2;分别比对照高17.1%、16.3%和9.5%;而施用蓝藻有机无机复混肥900kg/hm2和1200kg/hm2,韭菜的株高、叶宽和产量比对照分别提高了12.21%和13.32%;康乃馨产花量则以施肥量为1200kg/hm2的最高,产花量为4865枝/100m2,比对照高11.6%。韭菜和土壤样品中未检测出藻毒素含量,表明使用蓝藻有机无机复混肥对植物和土壤是安全的。利用水华蓝藻生产有机无机复混肥,既解决了富营养化湖泊大量暴发的水华蓝藻的巨大生物量处置难题,又达到了改善环境、增加了经济和社会效益。       

Semi-automatic algorithm for construction of the left ventricular area variation curve over a complete cardiac cycle

Background  

Two-dimensional echocardiography (2D-echo) allows the evaluation of cardiac structures and their movements. A wide range of clinical diagnoses are based on the performance of the left ventricle. The evaluation of myocardial function is typically performed by manual segmentation of the ventricular cavity in a series of dynamic images. This process is laborious and operator dependent. The automatic segmentation of the left ventricle in 4-chamber long-axis images during diastole is troublesome, because of the opening of the mitral valve.     

甘草酸的提取及其抑菌活性研究

本文对甘草酸的各种提取方法进行了对比研究。结果表明,超声提取与传统搅拌提取、酶解提取及反复冻溶提取相比,是较适于从甘草中快速高效提取甘草酸的新方法,其最优条件为用20倍量0.7%氨水超声处理2次,每次20min。此外,探讨了提取的甘草酸的体外抑菌活性,其对金黄色葡萄球菌,大肠埃希氏杆菌,绿脓杆菌和枯草杆菌的最小抑菌浓度(MIC)分别为:0.0625,0.125,0.25,0.03125mg/mL,表明其具有较好的抑菌活性。     

肺病患者呼吸道曲霉随机扩增多态性DNA基因型分析

目的：了解肺病患者呼吸道曲霉基因分型状况,寻找肺部曲霉感染的可能来源,为曲霉感染的防治提供参考。方法：选择有呼吸道症状的患者925例作受试对象,收集其鼻腔、咽部分泌物及支气管肺泡灌洗液（bronchoal veolar lavage fluid,BALF）进行真菌培养、鉴定及随机扩增多态性DNA（random amplified polymorphic DNA,RAPD）基因分型分析。结果：在烟曲霉、黄曲霉和黑曲霉的RAPD带形图中,扩增条带数在1～11条之间,片断大小在150bp～2kb不等,不同菌株之间扩增条带的数量和扩增片断的大小均存在差异,即使带型完全一致的菌株,有些片断也存在亮度的差异。对分离自同一患者呼吸道不同部位的同种曲霉种内相似性比较的结果显示：虽然来自同一患者呼吸道不同部位的同种曲霉的形态学特征相同,但是基因型特征却存在差异。结论：通过呼吸作用进入肺组织内部的曲霉与患者鼻腔定植的同种曲霉存在差异,提示肺部曲霉的感染可能来源于患者周围的环境中,有效的控制环境中曲霉的污染程度是预防肺部曲霉感染的最有利的手段。