Verschoor E J, van Vliet A L, Egberink H F, Hesselink W, Horzinek M C, de Ronde A
Institute of Infectious Diseases and Immunology, Veterinary Faculty, State University of Utrecht, The Netherlands.
J Clin Microbiol. 1993 Sep;31(9):2350-5. doi: 10.1128/jcm.31.9.2350-2355.1993.
The gag and env genes of the feline immunodeficiency virus strain UT113 were cloned into a baculovirus transfer vector. The recombinant plasmids were used to create recombinant baculoviruses that expressed either the gag or the env precursor protein in insect cells (Sf9 cells). Leader sequence cleavage occurred in Sf9 cells expressing the envelope precursor, but further processing was not observed. Crude lysates of insect cells infected with the wild-type baculovirus or with the recombinant viruses were used to develop an enzyme-linked immunosorbent assay for the detection of feline immunodeficiency virus-specific antibodies in cat sera. The assay showed a higher sensitivity and specificity than immunofluorescence and Western blotting (immunoblotting).
将猫免疫缺陷病毒UT113株的gag和env基因克隆到杆状病毒转移载体中。利用重组质粒构建重组杆状病毒,使其在昆虫细胞(Sf9细胞)中表达gag或env前体蛋白。在表达包膜前体的Sf9细胞中发生了前导序列切割,但未观察到进一步的加工过程。用野生型杆状病毒或重组病毒感染昆虫细胞的粗裂解物建立酶联免疫吸附测定法,用于检测猫血清中猫免疫缺陷病毒特异性抗体。该测定法比免疫荧光法和蛋白质印迹法(免疫印迹法)具有更高的灵敏度和特异性。