Preparation of Differentiated Recombinant Human β2-Microglobulin and Mouse β2-Microglobulin Proteins for its Detection in Class I HLA Chimeric Molecules
V. Karkischenko, V. Ezerskiy, E. Koloskova, M. S. Nesterov
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引用次数: 0
Abstract
Transgenic humanized animals are increasingly in demand for biomedical research and pharmacological testing. More and more lines of transgenic animals are being created, including those with knockout of their own genes. There is an urgent need for an evidence base for the integration of a transgene, its expression, determination of the knockout state of its own gene at the molecular genetic level, detection of translation of the target protein in different organs and tissues, proof for the absence of protein synthesis (or its non-functionality), the gene of which has been modified. This requires highly specific reagents, proteins and antibodies to them in particular, the vast majority of which are presented by foreign manufacturers. The task was set to identify mouse and human β2-microglobulin in protein fractions of organs and tissues of transgenic and knockout mice of several HLA lines created in recent years at the Scientific Center of Biomedical Technologies, Russia. At the first stage of our research, recombinant E. coli producing strains were obtained.
生物医学研究和药理测试对转基因人源化动物的需求越来越大。越来越多的转基因动物品系正在诞生,包括那些自身基因被敲除的转基因动物。转基因的整合、表达、在分子基因水平上确定自身基因的敲除状态、检测目标蛋白在不同器官和组织中的翻译、证明被修饰基因不合成蛋白质(或无功能),这些都急需一个证据基础。这需要高度特异性的试剂、蛋白质和抗体,尤其是这些试剂、蛋白质和抗体,而绝大多数试剂、蛋白质和抗体都是由国外制造商提供的。俄罗斯生物医学技术科学中心(Scientific Center of Biomedical Technologies)的任务是在近几年培育出的几种 HLA 系转基因小鼠和基因敲除小鼠的器官和组织的蛋白质部分中鉴定小鼠和人类的 β2-微球蛋白。在研究的第一阶段,我们获得了重组大肠杆菌生产菌株。