Russo S, Montermini L, Berkovitz-Siman-Tov R, Ponti W, Poli G
Institute of Microbiology and Immunology, Faculty of Veterinary Medicine, Milan, Italy.
FEBS Lett. 1998 Sep 25;436(1):11-6. doi: 10.1016/s0014-5793(98)00951-x.
The gp51-p30 glycoprotein constituting BLV envelope was expressed in Sf-21 insect cells by means of recombinant baculoviruses. Post-infection cell lysates were analyzed, in order to define the immunologic reactivity of recombinant products. Oligosaccharide chains, containing N-acetylglucosamine, mannose, galactose and sialic acid were found on recombinant gp51-p30. In order to investigate the timing of transcription and translation of the glycoprotein, kinetic assays were carried out on cell lysates and directly in situ on Sf-21 cells during the course of baculovirus infection. The use of different solubilizing reagents was also evaluated in order to rescue recombinant glycoprotein from its subcellular location.
构成牛白血病病毒(BLV)包膜的gp51-p30糖蛋白通过重组杆状病毒在Sf-21昆虫细胞中表达。对感染后细胞裂解物进行分析,以确定重组产物的免疫反应性。在重组gp51-p30上发现了含有N-乙酰葡糖胺、甘露糖、半乳糖和唾液酸的寡糖链。为了研究该糖蛋白转录和翻译的时间,在杆状病毒感染过程中对细胞裂解物以及直接在Sf-21细胞原位进行了动力学分析。还评估了使用不同的增溶试剂从亚细胞定位中挽救重组糖蛋白的情况。