蓖麻蚕不同组织器官中胰蛋白酶抑制剂的种类与分析

胰蛋白酶抑制剂(TI)是鳞翅目昆虫体内广泛存在的一种蛋白酶抑制剂,主要功能是抑制胰蛋白酶的降解。采用非变性聚丙烯酰胺凝胶电泳和胰蛋白酶抑制剂活性染色,对18个蓖麻蚕品种5龄第5天的血液及其它组织中TI种类的分布进行了调查。经碱性聚丙烯酰胺凝胶电泳,在血液中共检测到8种TIs,分别命名为ETI-1~ETI-8,品种间存在多态性分布;在头和脂肪体中分别检测到1种TI;在体壁中检测到3种TIs。经酸性聚丙烯酰胺凝胶电泳,在血液中共检测到4种TIs,分别命名为ETI-b1、ETI-b2、ETI-b3和ETI-b4,品种间种类分布无差异;在血液中检测到的TI包含了在头、脂肪体和体壁中检测到的TI种类。     

The Genetic Polymorphism Analysis in Tree Shrew(T.belangeri chinensis) based on Fluorescent Microsatellite

A total of 10 fluorescent labeling microsatellite markers detected by capillary electrophoresis were selected to analyze the genetic diversity of tree shrew(T.belangeri chinensis).Microsatellite Toolkit and PopGene softwares were used to calculate the genetic correlation indexes to assess the genetic diversity of the population.64 alleles with 6.4 alleles per locus were identified among 10 microsatellite loci.The average effective number of alleles, observed heterozygosity, expected heterozygosity and polymorphic information content were 3.7892, 0.6156, 0.6963, and 0.6367, respectively.Seven of all loci were high polymorphic, and three were moderate polymorphic.Five loci extremely significantly deviated from Hardy-Weinberg equilibrium(P<0.01).These novel polymorphic microsatellite markers indicated high genetic diversity, which will be used to evaluate genetic diversity in tree shrew.     

同位素稀释-高效液相色谱-线性离子阱质谱法测定总膳食乳类样品中8种三嗪类农药

本试验旨在建立用同位素稀释-高效液相色谱-串联线性离子阱质谱法（HPLC-LIT-MS）同时测定总膳食乳类样品中8种三嗪类农药的检测方法。在中国144个采样点采集总膳食乳类样品,在待测样品中加入氘代同位素内标,用乙腈经超声提取,凝胶渗透色谱（GPC）净化。以乙腈和含10 mmol/L甲酸铵和0.1%甲酸的水溶液为流动相,目标化合物经CAPCELL PAK CR 1：20色谱柱分离,采用电喷雾电离,选择反应监测（SRM）正离子模式监测二级子离子。以一一对应的氘代同位素为内标物,内标法定量,计算样品中三嗪类农药的含量。总膳食乳类样品中8种三嗪类农药的平均添加回收率为83.9%~103.3%;相对标准偏差为2.5%~9.1%;检出限（LODs）为0.003~0.300 μg/kg。结果表明,该方法精密度和准确度均满足总膳食乳类样品中8种三嗪类农药的膳食暴露评估要求。     

Seed mix type but not planting method or seed priming affect grassland restoration outcomes: a greenhouse trial

Commercial seed mixes and various mechanical planting methods have been implemented to overcome barriers to successful restoration of degraded grasslands but their effectiveness in aiding the restoration process has been understudied. Harvested seed must be cleaned of chaff to allow mechanical planting, which can exclude small seeds. We propose that fluid seed drilling techniques (suspending uncleaned harvested seed in a cellulose-based gel matrix, which may then be injected into the soil with a mechanical planter) may overcome these challenges. As a preliminary investigation of the proposed method we designed a greenhouse pot trial experiment aimed at quantifying restoration success through the measurement of seedling recruitment, biomass production and composition responses of commercial and harvested seed mixes under simulated fluid seed drilling and smoke water priming (collectively termed application method). These data were analysed using generalised linear modelling and multivariate analyses. Commercial mixed seed resulted in greater recruitment, whereas harvested seed produced the greatest biomass production. There was large multivariate separation between seed types and seed type × application method centroids for recruitment responses and only seed type for biomass responses. Application method had minimal effects on restoration measurements whilst seed type greatly affected these measurements. Careful selection of the restoration seed mix is advised.     

猪圆环病毒2型病毒样颗粒的纯化及鉴定

为获得较高纯度的猪圆环病毒2型（porcine circovirus type 2,PCV2） Cap蛋白的病毒样颗粒（virus-like particles,VLPs）并对其进行结构和功能研究,本试验以大肠杆菌表达系统表达了PCV2重组Cap （PCV2 rCap）蛋白,通过硫酸铵分级沉淀、Sephacryl S-300 HR凝胶过滤层析、Capto Q离子交换层析、CsCl密度梯度离心等方法对其进行了纯化。通过Western blotting对纯化所得蛋白进行免疫反应性鉴定,透射电镜（TEM）观察纯化蛋白的粒径及形态,高效液相尺寸排阻色谱（HPSEC）检测其组分分布。结果显示,纯化后PCV2 rCap蛋白经SDS-PAGE检测,可见28 ku的目的蛋白带,灰度扫描法检测其纯度为97.53%。Western blotting结果显示,该处的特异性蛋白条带有明显的免疫反应性;TEM检测其为粒径17.35~19.24 nm的规则球形颗粒,表明所获得的PCV2 rCap蛋白在表达时能在菌体内自我装配成VLPs,且在纯化过程中没有被破坏而解聚,仍保持天然状态;HPSEC检测纯化样品中VLPs含量为92.67%。本试验结果为进一步研究PCV2 VLPs的空间结构及其免疫保护机制提供了参考依据。     

Advance in serum proteomics research and technology

In recent years, great progress has been made in the theory and technology of proteomics. As embranchment of proteomics, serum proteomics has been payed more and more attention also. The advanced techniques have been applied in serum proteomics research, which promote the development of the methodology. A great advance has been made in serum proteomics research for seeking associated tumor markers, pharmacy and some non-cancer diseases.     

甘蔗与黑穗病菌互作的叶片差异蛋白分析

以高抗和高感黑穗病(Ustilago scitaminea Syd)的甘蔗(Saccharum complex)品种NCo376和Ya71-374为供试材料,在接种甘蔗黑穗病菌后分离叶片全蛋白,采用双向电泳技术,分析甘蔗黑穗病菌侵染后甘蔗叶片蛋白质表达谱的变化.研究结果表明,病菌侵染后,品种间、同一品种接种与对照间的叶片蛋白质表达谱均存在明显差异,部分表达上调,有些表达下调.对抗感病品种接种组中2个上调表达的蛋白质点的质谱进行进一步分析,其中一个是NBS类型的抗性蛋白,另一个是rieske Fe-S precursor protein,推测它们可能在甘蔗对黑穗病菌侵染的应答中起不同的重要作用.     

5-氮胞苷和利福平对丛枝病泡桐幼苗形态和蛋白质变化的协同影响

以感染丛枝病的豫杂一号泡桐幼苗为试验材料,通过不同浓度5-氮胞苷和利福平处理,研究了其幼苗形态和蛋白质的变化。结果表明,适宜浓度的利福平和5-氮胞苷共同使用可抑制泡桐丛枝病的发生。150μmol·L-15-氮胞苷+80 mg·L-1利福平可以抑制豫杂一号泡桐丛枝病症状。存在于健康豫杂一号幼苗中的蛋白质pI 6.8、m24ku而在病苗中观察不到。表明该蛋白质可能与豫杂一号泡桐丛枝病发生有一定关系。