以CP23重组蛋白为抗原建立检测微小隐孢子虫抗体间接ELISA方法的研究

摘要：以在E.coli高效表达的微小隐孢子虫子孢子表面抗原CP23为抗原，以辣根过氧化物酶(HRP)标记的山羊抗鼠IgG为二抗，建立了检测微小隐孢子虫抗体的间接ELISA方法。经检测筛选出最佳反应条件为1μg／孔纯化的E.coli表达的CP23抗原包被酶标板，用10％免血清进行封闭，以正常E.coli裂解上清液稀释待检血清。实验表明应用CP23重组蛋白作为诊断C.parvum抗原具有特异性高、抗原易纯化和成本低等特点。     

热带小奥德蘑深层发酵工艺研究

通过对热带小奥德蘑深层发酵培养基碳源、氮源和无机盐的筛选 ,利用正交试验得到了优化发酵培养基配方 ,在此基础上对发酵条件进行了探索。试验结果表明 ,热带小奥德蘑的适宜发酵培养基为 :葡萄糖 3%,蛋白胨 0 .4 %,KH2 PO40 .1%,MgSO4·7H2 O 0 .2 %,VB10 .0 1%。其适宜发酵温度为 2 8℃ ,起始pH5 .0～ 6.0 ,摇瓶装量 60mL 2 5 0mL ,接种量 10 %。     

J亚群禽白血病病毒与禽网状内皮增生症病毒共感染对肉鸡生长和免疫功能的抑制作用

1日龄网鸡感染J亚群禽白血病病毒(ALV-J)以及共感染禽网状内皮增生症病毒(REV)后,肉鸡生长发育明显受阻,体重增重明显下降(P＜0.05),法氏囊、胸腺明显萎缩(P＜0.05),在用新城疫疫苗免疫后,感染组血清中新城疫抗体效价显著低于对照组(P＜0.05);在ALV-J和REV共感染后,这种抑制作用更为明显(P＜0.01).ALV J单独感染后,鸡对传染性法氏囊病病毒(IBDV)弱毒疫苗免疫后的抗体反应与对照组没有明显差别,但ALV-J与REV的共感染可明显延缓鸡对IBDV弱毒疫苗免疫的抗体反应.     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

桃新品种‘乾兴银凤’

 ‘乾兴银凤’桃是从日本引进品种中选出的优良品种, 经品种比较试验和多点区域试验, 表现丰产, 抗逆, 不裂果等特性。     

鸡肉质鲜味特性相关基因的克隆及序列分析

本研究对泰和乌骨鸡、隐性白羽肉鸡腺苷琥珀酸裂解酶基因的cDNA进行了克隆,并对序列进行了分析,共发现有6处碱基突变249 G→A(TH)、717 C→G(TH)、985 A→G(TH)仅在泰和乌骨鸡中出现;992 T→C(RW)、1400 C→T(RW)仅在隐性白羽肉鸡中出现;而在泰和乌骨鸡和隐性白羽肉鸡中均检测到突变1179 A→C(TH,RW).其中985 A→G(TH)、1400C→T(RW)处突变导致两处氨基酸突变Thr→Ala(305)、Ala→Val(443),其它4处碱基突变均为同义突变.另外对几种脊椎动物Adsl基因核苷酸序列水平、蛋白质水平同源性进行了比较分析.     

用外翻肠囊法研究高钙条件下有机锰在肉仔鸡小肠中的吸收特点

本试验用外翻肠囊法研究高钙对肉仔鸡不同肠段(十二指肠、空肠和回肠)吸收不同形态锰的影响,以比较有机锰和无机锰在高钙条件下吸收特点的差异。采用单因子完全随机设计,培养液中添加的4种锰源分别为:硫酸锰、弱络合强度的蛋氨酸锰(Mn-MetE)、中等络合强度的氨基酸锰(Mn-AAA)和强络合强度的氨基酸锰(Mn-AAB)。为了扣除内源的影响,设置1个零添加锰水平处理。将40只31日龄AA肉公鸡随机分到以上5个组,每组8个重复,每个重复1只鸡,每只鸡的十二指肠、空肠和回肠分别作为相应肠段外翻肠囊的一个重复。结果表明:1)体外培养回肠肠囊对锰的吸收率显著高于十二指肠(P<0.01);2)回肠肠囊对强络合强度的Mn-AAB中锰的吸收率显著高于硫酸锰或中等络合强度的Mn-AAA(P<0.05)。本试验结果表明:在高钙条件下,络合形态的有机锰在体外培养的肉仔鸡肠囊中的吸收率显著高于无机形态的锰;强络合强度有机锰源中的锰能更强地抵抗小肠内的解离,比弱或中等络合强度的有机锰源更有利于锰的吸收。     

Progress in autoantibodies to platelet and idiopathic thrombocytopenic purpura

The onset of idiopathic thrombocytopenic purpura (ITP) is related to excessive destruction of platelet resulting from antiplatelet autoantibody. The immunity with ITP is imbalance, so the hypofunction of suppressor T cell (Ts) can't restrain B lymphocytes producing antibody, as a result, autoant ibody is produced, the clearance of platelet in blood circulation is accelerated. The different antibodies have different functions on ITP. The measuring of platelet antibody has its significance in diagnosis, therapeutic evaluat ion and expectation of prognosis.     

莠去津原药中莠去津及主要杂质的气质联用分析研究

本文通过采用气质联用(GC—MS)法对莠去津原药中莠去津及其它杂质进行分析鉴定，从而了解莠去津原药中的杂质情况和质量状况，用以指导工业生产。     

Characteristics for Practical Use of Attenuated Isolate UA-Fukushima of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis>

L₁₁A-Fukushima (L₁₁A-F) derived from attenuated isolate LuA of Tomato mosaic virus (ToMV) has the highest ability to cross protect against virulent ToMV among LuA and its derivatives and is stably inherited. Growth, yield, fruit quality and symptom attenuation of inoculated tomato plants did not differ significantly between L₁₁A-F and L₁₁A. The infectivity of progeny viruses in tomato infected with LuA-F was less than 4% of that with virulent ToMV. From these results, L₁₁A-F appears to possess the properties necessary for practical use. To manage L₁₁A-F strictly, a PCR-based assay to detect trace contamination of virulent ToMV in L₁₁A-F preparations was established. Received 10 June 2002/ Accepted in revised form 30 October 2002