Arnaoty Ahmed, Pitard Bruno, Bateau Benoit, Bigot Yves, Lecomte Thierry
GICC, UMR CNRS 6239, Université François Rabelais, UFR des Sciences et Technques, Tours, France.
Methods Mol Biol. 2012;859:293-305. doi: 10.1007/978-1-61779-603-6_17.
Molecular domestication of several DNA transposons has occurred during the evolution of the primate lineage, and has led to the emergence of at least 42 new genes known as neogenes. Because these genes are derived from transposons, they encode proteins that are related to certain recombinases, known as transposases. Consequently, they may make an important contribution to the genetic instability of some human cells. In order to investigate the role of these neogenes, we need to be able to study their expression as proteins, for example in tumours, which often provide good models of genetic instability. In order to perform such studies, polyclonal antibodies directed against the proteins expressed by neogenes are obtained using a recently developed new method of Nanospheres/DNA immunisation in laboratory mammals. In this chapter, we describe a fully integrated process of producing antibodies that consists of a series of steps starting with the preparation and synthetic formulation of plasmids encoding neogenes, and culminating in the final production and confirmation of the quality of these polyclonal antibodies.
在灵长类谱系的进化过程中，几种DNA转座子发生了分子驯化，并导致了至少42个新基因（即新起源基因）的出现。由于这些基因源自转座子，它们编码的蛋白质与某些重组酶（即转座酶）相关。因此，它们可能对某些人类细胞的遗传不稳定性有重要影响。为了研究这些新起源基因的作用，我们需要能够研究它们作为蛋白质的表达情况，例如在肿瘤中，肿瘤往往是遗传不稳定性的良好模型。为了进行此类研究，利用实验室哺乳动物中最近开发的纳米球/DNA免疫新方法，获得针对新起源基因表达的蛋白质的多克隆抗体。在本章中，我们描述了一个完整的抗体生产综合过程，该过程包括一系列步骤，从编码新起源基因的质粒的制备和合成配方开始，最终完成这些多克隆抗体的最终生产和质量确认。
