草莓PLDα基因cDNA克隆及反义植物表达载体构建

【目的】获得草莓磷脂酶Dα(PLDα)的HKD2功能区基因序列构建反义植物表达载体。研究PLDα基因对草莓果实耐贮性的影响。【方法】以草莓完熟果实为材料提取总RNA,利用RT-PCR技术扩增得到PLDα基因的cDNA片段,将其T-A克隆至pMD19-T simple vector上,测序正确后克隆载体经双酶切消化,目的片断反向插入到植物表达载体pBI121中。【结果】构建草莓PLDα基因的反义植物表达载体pBI121-PLD。【结论】通过冻融法将携带反义cDNA的植物表达载体质粒导入根癌农杆菌LBA44404,为进一步通过反义技术延长草莓果实贮藏的寿命奠定了基础。     

猪LXRα基因的克隆、序列分析及表达研究

采用 RT-PCR 结合克隆测序的方法,从猪的肝脏组织中克隆出了猪基因的 Liver X Receptors α cDNA 序列,编码区长度1344bp,编码447个氨基酸,基于Pfam数据库发现存在一个锌指蛋白和核受体的配体结合区;系统发育分析发现猪的LXRα所在的哺乳动物类群非常保守,与鸡和斑马鱼所构成的类群存在较大遗传距离;在大白猪和杂种野猪的脂肪组织中检测表明LXRα基因的表达随着月龄的增加也不断增加,并日趋稳定,而杂种野猪在9月龄开始高表达,12月龄最高。     

牛源金黄色葡萄球菌α溶血素单链抗体对小鼠乳腺的保护作用

为探究治疗奶牛乳腺炎的抗生素替代性生物制剂,研究将前期获得的特异性针对金黄色葡萄球菌毒力因子α溶血素的单链抗体scFvZZ107,在小鼠感染金黄色葡萄球菌乳腺炎模型中验证其保护作用。结果显示,3种不同剂量(5、10、15 μg)单链抗体处理组乳腺组织内细菌数量分别为25.42±1.3、3.33±0.53和4.8±0.78 lgcfu/mg,IL-1α的含量分别为49±7.91、31.37±2.26和57.34±4.48 pg/mL,IL-1β含量分别为40.91±7.39、47.81±4.78和39.2±2.28 pg/mL,IL-8的含量分别为48.64±1.86、41.65±3.07和45.74±1.56 pg/mL,与生理盐水处理组相比,单链抗体中、高剂量处理组乳腺内金黄色葡萄球菌检出量显著减少(P<0.05),且IL-1α、IL-1β与IL-8的含量均显著降低(P<0.05)。通过观察小鼠乳腺组织HE和TUNEL染色情况,与生理盐水处理组相比,scFv中、高剂量处理组乳腺组织更加完整,凋亡细胞的数量也更少,表明scFvZZ107对小鼠乳腺具有良好的保护作用。     

基于Alpha波的移动机器人控制系统

为了实现脑电信号对机器人的控制,根据脑电a 波的阻断现象,利用功率谱估计方法提取睁眼和闭眼时a 节律功率,并将闭眼时 a 节律功率的提升作为单次实验的触发,将其转换为外部机器人控制指令。该系统的工作模式为异步,可以任意选择何时启动和停止系统。实验表明,该系统对输入指令有较高的识别率,提供了一种新的控制机器人的方法。     

不同干扰梯度下尼泊尔森林树种多样性和优势种的变化(英文)

中度干扰可以增强多样性,因此干扰常作为一种管理手段。因而,了解森林干扰的频度和强度以及对林分结构、优势度以及森林多样性等的作用是非常重要的。本研究在尼泊尔设立25个1 ha样地,调查了不同干扰程度对以婆罗双树为优势种的5个林地生物多样性和优势度的影响。共记录67个树种,其中41个树种在轻度干扰的森林,10个在重度干扰的森林。林地之间α多样性差异显著。α多样性随干扰梯度线性降低,而优势度则呈线性增加。婆罗双树相对基面积随干扰强度而增加,重度干扰林下该树种重要值是轻度干扰林下的2倍。α多样性随婆罗叔树相对基面积的增加而以3倍的比率降低。在任意一对干扰林中,树种组成相似性均很低(相似性指数57%),说明β多样性较高。结论:婆罗双树林多样性随干扰强度增加而降低,但是优势度增加。建议控制优势种(婆罗双树)种群,来提高多样性,实现多功能利用的森林管理目标。图5表6参32。     

Protein display by bovine herpesvirus type 1 glycoprotein B

Glycoprotein B (gB) of bovine herpesvirus 1 (BHV-1), a major component of the viral envelope, is essential for membrane fusion during entry and cell-to-cell spread. It is cleaved in the trans-Golgi network by the proprotein convertase furin. Integration of the open reading frame (ORF) encoding a mutated gB with a second furin cleavage site and mature boIFN-α as intervening peptide between the amino-terminal (NH₂) and carboxy-terminal (COOH) gB subunits yielded recombinant BHV-1/gB2FuIFN-α which, unexpectedly, express gB with an enlarged NH₂-subunit of 90 kDa. Here we show that boIFN-α-specific antibodies bind to the 90 kDa gB subunit and efficiently neutralize BHV-1/gB2FuIN-α infectivity. We also show that inactivated BHV-1/gB2FuIN-α virions induce an antiviral state in cells incubated with UV-inactivated particles. These results demonstrate that the 90 kDa protein is a NH₂-subunit/boIFN-α fusion protein whose boIFN-α domain is biologically active. To verify that BHV-1 gB is suitable for the display of (glyco)proteins on the surface of virions we constructed BHV-1 recombinants expressing within gB the first 273 amino acids of the NH₂-subunit (HA1) of avian influenza haemagglutinin, either flanked by two furin cleavage sites or with only one cleavage site between a gB/NH₂_HA1 fusion protein and the COOH subunit. The resulting recombinant BHV-1/gB2FuHA1 expressed gB from which 55 kDa HA1 was excised and secreted. In contrast, gB from BHV-1/gB_NH₂HA1 infected cells retained HA1 as fusion protein with the NH₂-subunit. Immunoblotting and neutralization analyses revealed that HA1 is incorporated into the envelope BHV-1/gB/NH₂_HA1 particles and exposed to the exterior of virions. Thus, this novel approach enables display of polypeptides and (glyco)proteins of at least 273 amino acids on viral particles which is of particular interest for development of novel diagnostics and vaccines as well as for, e.g. gene therapy applications especially when biologically active ligands need to be presented.     

α抗胰蛋白酶研究进展

缺乏α抗胰蛋白酶可以引起肝炎、慢性肺阻塞等疾病.α抗胰蛋白酶的主要有抑制如凝血酶、纤溶酶、中性粒细胞弹性蛋白酶等多种丝氨酸的内切肽酶;抑制多形核白细胞(PMN细胞)的激活;清除细胞毒性物质,保护组织细胞并帮助其再生和修复的功能.本文就α抗胰蛋白酶理化性质、缺乏症极其抗病毒的治疗作用进行综述.     

天府肉鸭IFN-α基因的密码子使用法特征分析

密码子简并性特征造成了不同物种密码子使用的偏好性不同,这对外源基因的表达强度有着较大的影响。运用软件对选取的16种IFN-α和天府肉鸭IFN-α基因（EU925650）进行了密码子偏爱性分析。通过进化树的构建结果显示天府肉鸭IFN-α,与北京鸭、河南肉鸭以及绍兴鸭IFN-α的亲缘关系最为接近。通过CAI,ENC,GC3S等数据分析进行了密码子偏爱性特征的分析。结果显示,天府肉鸭IFN-α系统保守基因,与禽类IFN-α基因的密码子使用类似。     

河北和内蒙古马铃薯干腐病菌种类鉴定

为明确河北和内蒙古马铃薯干腐病菌种类和优势种群,从两省采集300块具干腐病症状的样品,通过分离培养、形态鉴定、TEF-1α序列分析及致病性测定等,对马铃薯干腐病菌种类进行了系统研究.结果表明,两省马铃薯干腐病共存在4种病原菌,即接骨木镰刀菌Fusarium sambucinum、锐顶镰刀菌F.acuminatum、尖孢镰刀菌F.oxysporum和芬芳镰刀菌F.redolens,分别为149、80、3、3株.其中,河北省的优势种群为接骨木镰刀菌和锐顶镰刀菌,其发生频率分别为50％和48.68％;内蒙古的优势种群为接骨木镰刀菌,发生频率为87.95％.     

Intrauterine infusion of BQ-610, an endothelin type A receptor antagonist, delays luteolysis in dairy heifers

Three separate in vivo experiments were conducted to evaluate the putative role of endothelin-1 (ET-1) during luteal regression in heifers. In Experiment 1, a single intraluteal injection of 500 μg BQ-610 [(N,N-hexamethylene) carbamoyl-Leu-d-Trp (CHO)-d-Trp], a highly specific endothelin A (ET_A) receptor antagonist, did not diminish the decline in plasma progesterone following a single exogenous injection of 25 mg prostaglandin F2 alpha (PGF₂) administered at midcycle of the estrous cycle. In Experiment 2, six intrauterine infusions of 500 μg BQ-610 given every 12 h on days 16–18 delayed spontaneous luteolysis, as evidenced by an extended elevation (P = 0.054) of plasma progesterone concentration. In Experiment 3, heifers were administered six intrauterine infusions of BQ-610 or saline on days 16–19, and peripheral blood samples were collected from day 11 to 16 (before infusion), hourly on days 16–19 (during infusion), and on days 20–25 (after infusion). BQ-610 treated heifers had markedly higher (P < 0.0001) levels of plasma progesterone compared with saline controls, and this effect was most notable during the infusion period (treatment by period interaction; P ≤ 0.05). Heifers infused with BQ-610 also had higher progesterone levels on day 21 (treatment by time interaction; P ≤ 0.05). Mean plasma concentrations of 13,14-dihydro-15-keto-PGF₂ (PGFM), the primary metabolite of PGF₂, were measured in the samples collected hourly and were not different (P ≥ 0.05) between treatments. These results indicate that the in vivo antagonism of the ET_A receptor can delay functional luteolysis, and supports the theory that ET-1 regulates luteal function in ruminants.