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

A. Schots  J. De Boer  A. Schouten  J. Roosien  J. F. Zil Verentant  H. Pomp  L. Bouwman-Smits  H. Overmars  F. J. Gommers  B. Visser  W. J. Stiekema  J. Bakker 《European journal of plant pathology / European Foundation for Plant Pathology》1992,98(2):183-191

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.  相似文献   

欢迎购阅下列新书     

YANG Yong-jian  ZHOU Xing-wen  ZHANG Xin  SU Xiao-hua  ZHU Jun 《园艺学报》2005,32(5):868-871

AIM: To investigate the contribution of angiotensin-converting enzyme inhibitor (ACEI) to the regulation of calpain system in infarcted myocardium. METHODS: Rat myocardial infarction (MI) model was established by permanent ligation of the left coronary artery. The treatment with the ACEI inhibitor rampril (1 mg·kg-1·d-1) was started 7 days prior to surgery. On day 1, 3, 7 and 14 after MI, protein levels of calpainⅠ, Ⅱ and calpastatin were determined in left ventricular free wall (LVFW), interventricular septum (IS) and right ventricule. RESULTS: CalpainⅠprotein level was increased in IS 14 d post MI, whereas the protein level of calpainⅡ was maximally increased in LVFW 3 d post MI. Rampril decreased protein up-regulation of calpainⅠ and Ⅱ, and reduced infarct size and interstitial fibrosis. Calpastatin protein expression was not affected by ACEI. CONCLUSIONS: CalpainⅠ is involved in cardiac remodelling in the late and calpainⅡ contributes to cardiac tissue damage in the early phase of MI. The heart protective effect of ACEI may be related to the inhibition of calpain system in the pathogenesis of myocardial infarction.  相似文献   

绵阳城市广场建设中的特色分析   总被引：1，自引：0，他引：1  

苏军 《西南园艺》2005,33(1):26-28

在调查的基础上，介绍了四川绵阳市的广场，并以铁牛广场为例分析其特色和不足。  相似文献   

野生朱红硫磺菌驯化栽培研究   总被引：2，自引：2，他引：2  

曾先富  廖志勇  张军 《中国食用菌》2005,24(6):18-20

营养试验、菌丝体培养和出菇温度试验、培养基酸碱度试验、出菇条件试验结果表明：朱红硫磺菌菌丝体适宜生长的温度范围为15-32℃，最适温度范围为22～28℃；子实体发生的最适温度为16～25℃，气温低于16℃，子实体色泽为淡橙色，16℃以上，子实体呈鲜橙红色；培养料适宜营养C／N为20-26／1；朱红硫磺菌菌丝体适宜酸碱度范围为pH3．0～8．0，最适酸碱度为pH4．0—6．5；朱红硫磺菌的产量与光照条件无关，但光照强度影响子实体的发生时间和色泽；朱红硫磺菌栽培原料广泛，凡栽培木腐菌的培养料均可用于栽培朱红硫磺菌。  相似文献   

组织培养条件下苹果Ty1-copia类逆转座子的转录活性     

孙俊  房经贵  陶建敏  蔡斌华  章镇 《果树学报》2005,22(5):441-445

研究了苹果基因组中Tyl—copia类逆转座子的转录活性、多样性及其甲基化水平。在培养基MS＋2，4-D2mg／L＋BA0．2mg／L＋NAA0．5mg／L上诱导了1个月的愈伤组织中用RT—PCR方法检测到了270bp的目的条带，而对照、继代组培苗和不含2，4-D的愈伤中均没有目的条带的扩增，说明一定浓度的2，4-D可以诱导苹果中Tyl—copia类逆转座子的表达活性。将目的条带回收、克隆和测序后进行分析：所克隆的21条序列虽都为Tyl-copia类逆转座子逆转录酶保守序列，但存在很大的异质性，21个克隆的核甘酸序列变化范围为196—290bp；翻译成氨基酸后存在缺失、插入、移框和终止密码子突变，7个克隆在氨基酸保守序列‘SLYGLKQ’处有1—2氨基酸位点的突变，说明这些序列对应的逆转录酶多数可能已失去了其原有功能。将21条序列进行聚类分析，除ART20外，其它克隆可聚为3类，并分别与GenBank上某些物种的相应序列同源性很高，表明在不同物种间可能存在逆转座子的横向传递作用。甲基化试验表明苹果基因组内Tyl-copia类逆转座子甲基化水平高。这些都表明Tyl-copia类逆转座子对苹果基因型的多样性及基因组的进化具有重要意义。  相似文献   

柑橘裂皮病类病毒的一步RT-PCR法检测及其在甜橙体内的分布   总被引：3，自引：0，他引：3  

彭军  周常勇  唐科志  谭万忠 《果树学报》2005,22(6):744-747

柑橘裂皮病是由柑橘裂皮病类病毒(Citrusexocortisviroid,CEVd)引起的一种重要的柑橘病害。分别采用快速微量核酸提取法和SDS-KAc抽提法在表现症状的Etrog香橼中提取核酸,采用一步RT-PCR法对CEVd进行检测,结果显示SDS-KAc法可以消除带毒样品中非特异性条带。从嫁接接毒的铜水72-1锦橙植株的接穗部取嫩叶、老叶、皮,从砧木部茎杆上取老皮以及根皮分别提取总核酸进行RT-PCR检测,分析CEVd在甜橙体内的分布情况。初步检测结果表明在接穗的嫩叶、老叶、皮,砧木部茎杆的老皮和根皮上都可以检测到CEVd,以接穗部叶和皮检出稳定性高。  相似文献   

母猪卧向行为学的观察     

邓红雨  范佳英  包军 《家畜生态》2005,(5)

本文通过2个试验对母猪在分娩圈和分娩栏中的卧向行为进行了观察,试验一中, 选择10头大白(Yorkshire)经产母猪作为观察对象,试验二中分别选择10头大白(Yorkshire) 经产母猪和10头长白(Landrace)经产母猪作为观察对象。观察采用瞬时记录方法,每周观察 三次,隔日观察,每天上午、下午各观察一次,每次3 h,每次观察间隔5 min。观察中发现,母猪 卧向以向外为主。分娩栏(四周是栏杆)中的母猪以选择卧向外(向北)为主,其次是卧向内(向 南),卧向左和卧向右最少且差异不显著,上栏前和下栏后差异不显著。分娩圈(四周是墙壁)中 母猪以选择卧向外为主,卧向内最少,卧向左和卧向右差异不显著。长白母猪比大白母猪选择 卧向外的多,妊娠阶段比哺乳阶段选择卧向外的多。  相似文献   

菌根与植被恢复   总被引：5，自引：1，他引：5  

林晓民  李振岐  韩德俊  侯军 《干旱地区农业研究》2003,21(2):167-170

菌根不仅可促进植物的营养吸收、生长发育、抗病、抗逆,而且在保持良好的土壤结构、控制水土流失方面具有直接的作用。在我国西北许多地区,菌根菌与植物间的共生关系已被中断,要恢复该地区的植被,治本的方法应是重建植物与菌根菌的共生关系,形成健康的生态系统。文中还提出了菌根技术在植被恢复中的应用策略。  相似文献   

Selection and amplification of the liver stem cell subset from rat bone marrow cells with a medium containing cholestatic serum in vitro     

CAI Yun-feng  MIN Jun  FANG Tian-ling  CHU Zhong-hua  DENG Xiao-geng  HE Jing-song  CHEN Ji-sheng 《园艺学报》2003,19(8):1029-1033

AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro. METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy.  相似文献   

Effect of mutant β-catenin gene on the proliferation of hepatocytes     

SHANG Xian-zhang  MIN Jun  CHU Zhong-hua  CHEN Ji-sheng 《园艺学报》2003,19(12):1601-1605

AIM:The β-catenin is a key molecule in the Wnt signal pathway, which plays a critical role in normal development and tumorigenesis. However, the mechanisms of the β-catenin on the cell growth control are still not completely defined. The aim of this study was to test the hypothesis that the mutant β-catenin may regulate the hepatocyte proliferation. METHODS: The immortalized murine hepatocyte cell line, AML12, was used for this study. A plasmid that contain mutant β-catenin S33Y was transfected into the AML12 cells and a stable cell line AML12S33Y was established. The cell growth property of this cell line and the parental cell were compared by flow cytometry analysis and direct cell count. The cells were also tested for the ability to form soft agar colonies, and the ability to form tumors in the severe immune deficient mice (SCID). RESULTS:1. The mutant β-catenin containing cell line AML12S33Y has higher proliferating index compared with the parental AML12 cells (P<0.01), suggesting that mutant β-catenin promotes cell growth. 2. The mutant β-catenin cells formed small colonies in soft agar after 4 weeks of culture, but did not generate tumor in SCID mice. CONCLUSION:The mutant β-catenin promotes liver cell growth.  相似文献