生长抑素(SS)与硫氧还原蛋白(Thioredoxin)相融合生成的2种基因工程化融合蛋白SS-N/X和SS-N/S的表达特性

对基因工程化生长抑素（somatostatin,SS)-硫氧还原蛋白（Thioredoxin)融合蛋白SS－N／X和SS－N／S的表达产量、水溶性、Triton X-100对SS融合蛋白水溶性的影响、包涵体的溶解和复性特点以及SS的抗原性与免疫原性等特性进行了研究。结果显示，在30℃的低温下用IPTG诱导表达，SS－N／X融合蛋白主要以可溶性形式存在，占75．8％，色涵体仅占24．2％；而SS－N／S融合蛋白则主要以包涵体的形式存在，占81．1％，可溶性蛋白仅占18．9％；32C载体（pEG-32C)蛋白则居二者之间，可溶性蛋白与包涵体大约等比例出现。SS－N／S在低温下诱导可超量表达SS融合蛋白，其表达量约占菌体总蛋白的40．94％。0．5％ Triton X-100对融合蛋白的不溶性有非常显著的影响，几乎可使所有的SS－N／X融合蛋白和大部分SS－N／S融合蛋白及32C硫氧还原蛋白变成水溶性的。32C硫氧还原蛋白包涵体在尿素中的溶解度明显高于SS－N／S包涵体，2mol/L尿素可使近一半的32C硫氧还原蛋白包涵体溶解，而SS－N／S包涵体只有27．5％溶解；但在5mol/L尿素时，两者均可基本全部变为可溶性的。在SS融合蛋白包涵体溶解时加入还原剂二硫苏糖醇，可明显降低SS的抗原性。SS融合蛋白（水溶性融合蛋白和复性后的包涵体）具有良好的SS抗原性和免疫原性。由SS融合蛋白与弗氏不完全佐剂制成试验用疫苗，免疫BALB／c小鼠、仔猪和羔羊等动物，可显著提高免疫动物的增重。     

Using laser to measure stem thickness and cut weed stems

Summary Stem thickness of the weed Solanum nigrum and the crop sugarbeet was determined with a He–Ne laser using a novel non‐destructive technique measuring stem shadow. Thereafter, the stems were cut close to the soil surface with a CO₂ laser. Treatments were carried out on pot plants, grown in the greenhouse, at two different growth stages, and plant dry matter was measured 2–5 weeks after treatment. The relationship between plant dry weight and laser energy was analysed using two different non‐linear dose–response regression models; one model included stem thickness as a variable, the other did not. A binary model was also tested. The non‐linear model incorporating stem thickness described the data best, indicating that it would be possible to optimize laser cutting by measuring stem thickness before cutting. The general tendency was that more energy was needed the thicker the stem. Energy uses on a field scale are discussed.     

家鸡Leptin成熟肽cDNA的克隆、重组蛋白表达及纯化

从 18周龄鸡卵巢组织中抽提总 RNA,使用六聚体随机引物反转录后 ,用鸡 L eptin(瘦素 )特异性引物扩增出鸡L eptin编码区第 5 2～ 4 6 0 bp的长度为 4 0 9bp的 c DNA片段。根据鸡 L eptin的 3′端第 4 6 0～ 4 92 bp序列设计了 3条部分相互重叠并且顺序串联延伸的下游反义引物 ,并在最后 1条引物 3′端连接上 Eco R 切点 ;在以上用于反转录扩增的 5′端引物的 5′端连接一 Bam H 切点。用该 5′端引物分别与 3个 3′端引物配对 ,利用反转录扩增出的 4 0 9bp的L eptin c DNA片段作为第 1模板进行扩增 ,扩增产物再作为模板与下一引物对再次扩增。经 3次扩增得到编码鸡 L ep-tin成熟肽的全长 4 5 1bp的 c DNA序列。将该 4 5 1bp的 L eptin c DNA序列经 Bam H 和 Eco R 双酶切后 ,克隆入表达质粒 p RSET A的 Bam H 和 Eco R 两酶切位点之间 ,构建成表达质粒 p L ep- SCAU。转化有重组表达质粒 p L ep-SCAU的大肠杆菌 BL 2 1(DE3)在 L B培养基中培养后 ,经 IPTG诱导表达出相对分子质量为 2 0 10 0的鸡 L eptin融合蛋白和少量 4 0 2 0 0的 L eptin融合蛋白。L eptin融合蛋白的表达在 IPTG浓度为 0 .0 5 mmol/ L 时达到最高 ,占总菌体蛋白的 32 .6 %。用 Ni- NTA凝胶从 7L 发酵培养菌裂解液中纯化出 180 m g左右     

Visualization of the effect of a surfactant on the uptake of xenobiotics into plant foliage by confocal laser scanning microscopy

A radiolabelling method is generally used to determine the foliar uptake of xenobiotics. This technique cannot provide any information on the localization of chemicals inside leaf tissues. The influence of an alcohol ethoxylate surfactant on the uptake of three fluorescent dyes, difluorofluorescein (hydrophilic), rhodamine B (moderately lipophilic) and a naphthalimide dye (lipophilic), into the leaves of three contrasting species, bean (Vicia faba), wheat (Triticum aestivum) and cabbage (Brassica oleracea), at 16 h after treatment was measured using a conventional wash‐off method and also visualized in vivo by confocal laser scanning microscopy (CLSM). Whereas the lipophilic dyes showed greater intrinsic uptake than the hydrophilic one, the enhancing effect of the surfactant on uptake was more pronounced for the latter. CLSM revealed that the presence of the surfactant increased the transport of difluorofluorescein into the epidermal cells of bean and wheat leaves, but not cabbage leaves. Rhodamine B showed greatest transcuticular diffusion in all three species, but most of the dye moved into the vacuole of the epidermal cells. The naphthalimide dye was strongly retained by the wax–cuticle layer of all species, even in the presence of the surfactant. CLSM has proven to be an attractive tool for studying xenobiotic diffusion. The results obtained using fluorescent dyes are believed to be applicable to the foliar uptake of herbicides.     

脉冲激光轰击法连续制备纳米金掺杂YBCO

利用超声法获得钇钡铜氧（YBCO）和乙醇混合溶液,采用聚焦脉冲激光轰击浸于流动无水乙醇中的金靶,连续制备得到纳米金和乙醇混合溶胶.将纳米金和乙醇混合溶胶直接滴入接收器中的YBCO和乙醇混合溶液中,经干燥获得纳米金掺杂的YBCO材料.结果发现,纳米金未明显改变YBCO的吸放热现象,对样品的组成影响不大;掺杂0.01%、0.05%重量比例的纳米金后YBCO的结晶度和超导转变温度Tc均有所提高.     

三种融合标签对大熊猫轮状病毒结构蛋白VP6-VP7可溶性表达的影响

大熊猫轮状病毒(giant panda rotavirus,GPRV)是引起幼龄大熊猫腹泻的主要病原,对圈养大熊猫产生了较大的危害。轮状病毒结构蛋白VP6是一种载体蛋白,可介导黏膜免疫反应。VP7是轮状病毒结构蛋白中主要的中和抗原。因此,VP6-VP7的融合表达作为候选抗原对该病的防治具有重要的意义。传统大肠埃希菌原核表达存在表达量低、可溶性差以及纯度低等弊端。本研究使用醛缩酶(EDA)、谷胱甘肽S-转移酶(GST)、麦芽糖结合蛋白(MBP)3种融合标签,以实现获得表达量高和纯度高的GPRV-VP6-VP7重组表达蛋白。将扩增的VP6、VP7基因片段利用同源重组酶构建到含3种融合标签的表达载体pET21b上,将重组质粒转化至大肠埃希菌Rosetta(DE3)感受态细胞中进行低温诱导表达。用Ni-柱亲和层析法纯化目的蛋白,SDS-PAGE和Image J分析蛋白表达量和可溶性,Western blot分析得到表达的重组表达蛋白正确且具有蛋白活性。实验结果证明,EDA标签能显著促进VP6-VP7蛋白的原核可溶性表达,提高VP6-VP7蛋白表达量。     

Assessment of Vein Diameter and Body Surface Temperature after High-Intensity Laser Therapy (HILT) on the Tarsal Joint in Healthy Horses

The aim of the study was to assess the influence of high-intensity laser therapy (HILT) on vein diameter and on body surface temperature changes in the tarsal joint area in a group of clinically healthy racehorses. The hypothesis of the study was that HILT increases vein diameter and body surface temperature in healthy tissue. The study involved 16 Thoroughbreds being subjected to ultrasonographic examination to assess changes of diameter of the cranial branch of the medial saphenous vein and thermographic examination to indicate temperature change of the dorsal surface of the tarsal joint, just before and immediately after HILT treatment. Vein diameter and mean surface temperature of the tarsal joint significantly increased after HILT treatment. In addition, the study was the first to describe the photothermal effect of HILT in healthy horses. More studies are necessary to specify the parameters of the procedure, that is, wavelength, energy density, or time of the procedure, depending on the patient’s individual characteristics and type of tissue.     

被征地农民的群体分化与社区融合——以重庆为例

在对重庆市进行实地调研的基础上，从群体、社区以及社会工作者3个方面来探寻影响被征地农民社区融合的原因，在分析不同类型被征地农民社区融合的需求的基础上，探讨不同层次的社会工作介入被征地农民社区融合的路径。