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 (VL) and heavy chain (VH) 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
1. An experiment was done under commercial conditions to investigate the physiological effects of isolating broody turkey hens, for 72 h, in sand and wire floored pens on the third, 10th and 16th weeks of production.
2. Hens identified as broody and removed from the flock had higher plasma prolactin concentrations than the laying hens at each of the three experimental stages.
3. Confinement in sand and wire floored pens, induced a decline in plasma prolactin concentrations. This decline probably impeded immediate development of broody behaviour. Alternately, levels of prolactin higher than those of laying hens were again measured 7 and 14 days after treatment during third week but not after the 10 th and 16th week of production.
4. Confinement did not induce consistent changes in luteinising hormone (LH) and progesterone concentrations from one period to an other.
5. An increase in the plasma concentration of D‐(/?)‐hydroxybutyrate was observed in the hens which had an egg present in the oviduct on day 2, 3 and/or 4 of the treatment. Subsequently, a decrease in ovulation rate was observed in the hens with higher concentrations of D‐(β)‐hydroxybutyrate while under treatment, during the 10th week of production.
6. These data confirm that the effectiveness of the traditional methods for broodiness prevention under commercial conditions is related to the induction of a decrease of prolactin. 相似文献
