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    • 5. 发明授权
    • Nucleic acids obtained from the envelope coding region of feline immunodeficiency virus molecular clone designated JSY3
    • 从猫免疫缺陷病毒分子克隆的包膜编码区获得的核酸为JSY3
    • US06331616B1
    • 2001-12-18
    • US08811682
    • 1997-03-05
    • Wayne TompkinsMary TompkinsJoo-Sung Yang
    • Wayne TompkinsMary TompkinsJoo-Sung Yang
    • C07H2104
    • C07K14/005A01K67/027A61K39/00C12N2740/15022
    • A full-length feline immunodeficiency virus NCSU1 (FIV-NCSU1) genome (JSY3) was cloned directly from FIV-NCSU1-infected feline CD4+ lymphocyte (FCD4E) genomic DNA and identified by PCR amplification with 5′ long terminal repeat (LTR), gag, env, and 3′ LTR primer sets. Cell-free JSY3 virus was cytopathogenic for FCD4E lymphocytes but did not infect CrFK cells in vitro. To determine in vivo infectivity and pathogenesis, six young adult specific-pathogen-free cats were inoculated with cell-free JSY3 virus. Provirus was detected at 2 weeks postinfection (p.i.) and was still detectable at 25 weeks p.i. as determined by gag region PCR-Southern blot analysis of peripheral blood mononuclear cell lysates. Infectious virus was recovered from peripheral blood mononuclear cells at 6 and 25 weeks p.i., and an antibody response to FIV was detected by 4 weeks. In the acute phase of infection, JSY3 provirus was found only in the CD4+ lymphocyte subset; however, by 14 weeks p.i., the greatest provirus burden was detected in B lymphocytes. All six cats were panlymphopenic at 2 weeks p.i., CD4+/CD8+ ratios were inverted by 6 weeks p.i., and five of the six cats developed lymphadenopathy by 10 weeks p.i. The claimed invention is directed toward isolated nucleic acids corresponding to the full-length JSY3 env coding region (nt. 6269-8824) and an env fragment comprising the transmembrane spanning domain (nt. 8339-8374). These nucleic acids will prove useful, inter alia, as molecular probes for FIV-specific sequences and in the generation of FIV-specific antigens and immunological reagents.
    • 从FIV-NCSU1感染的猫CD4 +淋巴细胞(FCD4E)基因组DNA中直接克隆全长猫免疫缺陷病毒NCSU1(FIV-NCSU1)基因组(JSY3),并通过5'长末端重复(LTR),gag ,env和3'LTR引物组。 无细胞的JSY3病毒对于FCD4E淋巴细胞是细胞致病性的,但在体外未感染CrFK细胞。 为了确定体内感染和发病机制,将6只年轻成年无特异病原体的猫用无细胞的JSY3病毒接种。 在感染后2周检测到病毒(p.i.),并且在25周p.i仍然可检测到。 通过gag区PCR确定外周血单核细胞裂解物的PCR- DNA印迹分析。 在6周和25周时从外周血单核细胞中回收感染性病毒,4周后检测到对FIV的抗体反应。 在感染的急性期,仅在CD4 +淋巴细胞亚群中发现JSY3原病毒; 然而,在14周的时间内,B淋巴细胞中检测到最大的病毒负担。 所有六只猫在2周时均为泛淋巴细胞,CD4 + / CD8 +比例在6周时反转,六只猫中有五只发生10周p.i的淋巴结肿大。 所要求保护的发明涉及对应于全长JSY3环编码区(nt.6269-8824)的分离的核酸和包含跨膜跨膜结构域(nt.8339-8374)的env片段。 这些核酸尤其可用作FIV特异性序列的分子探针和FIV特异性抗原和免疫试剂的产生。