中文摘要：本发明公开了一种口蹄疫疫苗的微环DNA载体。微环DNA载体的转基因表达盒包括：一个真核翻译起始序列；一个编码口蹄疫病毒衣壳蛋白前体的突变氨基酸序列，该序列至少包含一个突变点并产生限制性酶切位点；一个编码蛋白酶的氨基酸序列，该蛋白酶可以将口蹄疫病毒衣壳蛋白前体剪接为多种口蹄疫病毒衣壳蛋白和一个可调控蛋白酶表达的翻译调控元件。该微环DNA载体可直接转染哺乳动物宿主细胞，当转染至宿主细胞后，能够产生病毒样颗粒，从而刺激哺乳动物的免疫系统建立起抗口蹄疫病毒的免疫反应。
外文摘要：

This application is directed generally to minicircle DNA vectors for the vaccination of foot-and-mouth disease (FMD). The transgene expression cassette in the minicircle DNA vector includes: a eukaryotic translation initiation nucleotide sequence, a mutant nucleotide sequence that encodes a foot-and-mouth disease virus (FMDV) capsid polyprotein precursor that contains at least one mutation to eliminate a restriction enzyme recognition site, a nucleotide sequence that encodes a protease that cleaves the FMDV capsid polyprotein precursor into a plurality of FMDV capsid proteins and a translational regulatory element to regulate the expression of the protease. The minicircle DNA vectors can be transfected directly into the cell of a mammalian host. When transfected into the mammalian host cell, virus-like particles can be produced intrinsically to stimulate the mammalian host's immune system to develop adaptive immunity toward foot-and-mouth disease.
NOVELTY - A mutant nucleotide sequence that encodes a footAnd-mouth disease virus (FMDV) capsid polyprotein precursor derived from an FMDV, the mutant nucleotide sequence comprising a mutation that removed a restriction enzyme recognition site from a nucleotide sequence from which the mutant nucleotide sequence was formed, is new.
USE - The mutant nucleotide sequence or vector is useful for vaccinating a mammal against an FMDV; and determining whether a mammal is vaccinated against or infected with FMDV (all claimed).
ADVANTAGE - A minicircle DNA vector vaccine platform for FMD is provided.

主权项：

DETAILED DESCRIPTION - INDEPENDENT CLAIMS are included for:

(1) a vector comprising a mutant nucleotide sequence defined above;

(2) a transformed host cell comprising a mutant nucleotide sequence defined above;

(3) a virus-like particle (VLP) comprising a polypeptide produced from expression of a vector comprising a mutant nucleotide ! sequence defined above;

(4) vaccinating a mammal against an FMDV by administering a vector comprising a mutant nucleotide sequence defined above which when expressed in a host cell of the mammal induces production of footAnd-mouth disease VLP by the host cell; and

(5) determining whether a mammal is vaccinated against or infected with FMDV by detecting an antibody's presence in a sample from the mammal; and detecting an other antibody's presence or absence in the sample, the absence of the other antibody indicates vaccination of the mammal with a vector comprising a mutant nucleotide sequence defined above which when expressed in a host cell of the mammal induces production of FMDV VLPs.

申请号：US201514962272
公开/公告号：US2017158739A1
申请日：2015-12-08
公开/公告日：2017-06-08
申请/专利权人：US DEPT HOMELAND SECURITY (USGO-C)
发明/设计人：PUCKETTE, M;RASMUSSEN, M;NEILAN, J
分类号：C07K14/005; G01N33/56983; A61K39/00; C12N2770/32122; C12N2770/32123; C12N2770/32134; C12N2770/32143; G01N2333/09; G01N2469/20; G01N2800/26
主分类号：C07K14/005; G01N33/569
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