Rola-Łuszczak Marzena, Grabowska Agnieszka, Szewczyk Bogusław, Kuźmak Jacek
Department of Biochemistry, National Veterinary Research Institute, 24-100 Puławy, Puławy Poland.
Laboratory for Recombinant Vaccines, Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk, 80-309 Gdańsk, Gdańsk, Poland.
J Vet Res. 2019 Mar 22;63(1):1-6. doi: 10.2478/jvetres-2019-0020. eCollection 2019 Mar.
Field isolates of bovine leukaemia virus (BLV) show the presence of a few amino acid substitutions in major conformational G and H epitopes on surface glycoprotein gp51. Potentially, these substitutions can affect the 3D structure of these epitopes leading to their diminished immunoreactivity. The aim of this study was to express three gp51 glycoproteins carrying mutated epitopes as recombinant baculovirus proteins in insect cells to test their immunoreactivity with bovine sera.
gene chimeras encoding mutated epitopes G and H in the backbone of BLV FLK strain were constructed, cloned into pFastBac1 vector, and expressed in baculovirus.
The presence of recombinant gp51 protein in Sf9 insect cells was confirmed using monoclonal antibodies. ELISA tests were developed to check the immunoreactivity of recombinant protein with bovine sera.
Recombinant gp51 proteins with altered G and H epitopes can be used for further studies to analyse the serological response of bovine sera towards BLV antigenic variants.
牛白血病病毒（BLV）的野外分离株显示，表面糖蛋白gp51上的主要构象G和H表位存在一些氨基酸替换。这些替换可能会影响这些表位的三维结构，导致其免疫反应性降低。本研究的目的是在昆虫细胞中表达三种携带突变表位的gp51糖蛋白作为重组杆状病毒蛋白，以测试它们与牛血清的免疫反应性。
构建了在BLV FLK株骨架中编码突变表位G和H的基因嵌合体，克隆到pFastBac1载体中，并在杆状病毒中表达。
使用单克隆抗体确认了Sf9昆虫细胞中重组gp51蛋白的存在。开发了ELISA试验来检测重组蛋白与牛血清的免疫反应性。
具有改变的G和H表位的重组gp51蛋白可用于进一步研究，以分析牛血清对BLV抗原变体的血清学反应。
