Temperature-sensitive attenuated FMDV strains, construction method and application thereof
AssigneeHarbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences
InventorsLi Yu, Decheng Yang, Haiwei Wang, Guohui Zhou
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Abstract
A temperature-sensitive attenuated FMDV strain, construction method and applications thereof. The construction method of the temperature-sensitive attenuated FMDV strain is as follows. Mutating a cytosine on K region loop of IRES domain 4 of an FMDV genome to a guanine or an adenine to obtain the temperature-sensitive attenuated FMDV strain, or replacing a K region of IRES domain 4 of an FMDV genome with a K region of IRES domain 4 of a bovine rhinovirus genome to obtain the temperature-sensitive attenuated FMDV strain.
Claims9
A construction method of a temperature-sensitive attenuated FMDV strain, comprising:
- constructing a FMDV full-length cDNA infectious cloning plasmid, performing a site-directed mutagenesis to an IRES region of the FMDV full-length cDNA infectious cloning plasmid, obtaining a FMDV genomic RNA by in vitro transcription, and transfecting the FMDV genomic RNA into cells and culturing the cells containing the FMDV genomic RNA at temperatures to rescue the temperature-sensitive attenuated FMDV strain;
- wherein a cytosine at 351-site on K region loop of IRES domain 4 of the FMDV genomic RNA obtained by the in vitro transcription is mutated to a guanine or an adenine during the site-directed mutagenesis, a base sequence of the K region loop after mutation is 351GUUUAA356 or 351AUUUAA356.
A construction method of a temperature-sensitive attenuated FMDV strain, comprising:
- constructing a FMDV full-length cDNA infectious cloning plasmid, obtaining a FMDV genomic RNA by in vitro transcription, and transfecting the FMDV genomic RNA into cells to rescue the temperature-sensitive attenuated FMDV strain;
- wherein an IRES domain 4 of the FMDV genomic RNA obtained by the in vitro transcription is replaced with an IRES domain 4 of a bovine rhinovirus genomic RNA.
A chimeric IRES sequence obtained by replacing an IRES domain 4 of a foot-and-mouth disease virus genomic RNA with an IRES domain 4 of a bovine rhinovirus genomic RNA, wherein the chimeric IRES sequence has the nucleotide sequence of SEQ ID No:
- 2.
A construction method of a temperature-sensitive attenuated FMDV strain, comprising:
- constructing a FMDV full-length cDNA infectious cloning plasmid, obtaining a FMDV genomic RNA by in vitro transcription, and transfecting the FMDV genomic RNA into cells to rescue the temperature-sensitive attenuated FMDV strain;
- wherein a K region of an IRES domain 4 of the FMDV genomic RNA obtained by the in vitro transcription is replaced with a K region of IRES domain 4 of a bovine rhinovirus genomic RNA.
A chimeric IRES sequence obtained by replacing a subdomain K of FMDV IRES domain 4 with a subdomain K of IRES domain 4 of a bovine rhinovirus, wherein the chimeric IRES sequence has the nucleotide sequence of SEQ ID No:
- 3.
Dependencies are inferred from the claim text.