The present invention relates to a nucleotide sequence as shown in SEQ ID NO: 1 for encoding a Marburg virus envelope glycoprotein, and to a human replication-deficient recombinant adenovirus capable of expressing the nucleotide sequence and a preparation method therefor, as well as an application thereof in the preparation of a vaccine against Marburg virus disease. The vaccine uses an E1 and E3 deleted replication-deficient human type-5 adenovirus as a vector, and HEK293 cells integrating an adenovirus E1 gene as a packaging cell line, and a protective antigen gene carried is a codon-optimized Marburg virus Angola strain envelope glycoprotein gene. After codon optimization of the envelope glycoprotein gene, significant expression of envelope glycoprotein can be detected in transfected cells.

The present invention relates to a nucleotide sequence as shown in SEQ ID NO: 1 for encoding a Marburg virus envelope glycoprotein, and to a human replication-deficient recombinant adenovirus capable of expressing the nucleotide sequence and a preparation method therefor, as well as an application thereof in the preparation of a vaccine against Marburg virus disease. The vaccine uses an E1 and E3 deleted replication-deficient human type-5 adenovirus as a vector, and HEK293 cells integrating an adenovirus E1 gene as a packaging cell line, and a protective antigen gene carried is a codon-optimized Marburg virus Angola strain envelope glycoprotein gene. After codon optimization of the envelope glycoprotein gene, significant expression of envelope glycoprotein can be detected in transfected cells.

The present invention includes methods of making, and compositions comprising, a uniformly lethal filovirus for outbred small mammals by mutation of the viral genome through serial passages in a small mammal, the method comprising the steps of: obtaining a filovirus strain from a human subject; passing the filovirus strain one or more times by intramuscular injection of one or more filovirus infected tissues into an inbred small mammal until uniform lethality is obtained; passing the filovirus strain in one or more human cell lines; passing the filovirus strain one or more times by intraperitoneal injection of one or more filovirus infected tissues into an outbred small mammal until uniform lethality is obtained; and isolating the uniformly lethal filovirus obtained thereby.

The present invention includes methods of making, and compositions comprising, a uniformly lethal filovirus for outbred small mammals by mutation of the viral genome through serial passages in a small mammal, the method comprising the steps of: obtaining a filovirus strain from a human subject; passing the filovirus strain one or more times by intramuscular injection of one or more filovirus infected tissues into an inbred small mammal until uniform lethality is obtained; passing the filovirus strain in one or more human cell lines; passing the filovirus strain one or more times by intraperitoneal injection of one or more filovirus infected tissues into an outbred small mammal until uniform lethality is obtained; and isolating the uniformly lethal filovirus obtained thereby.