Title:
REVERSE GENETICS SYSTEM FOR FELINE MORBILLIVIRUS
Document Type and Number:
WIPO Patent Application WO/2023/230598
Kind Code:
A3
Abstract:
Disclosed are methods of producing recombinant feline morbillivirus (FeMV) using reverse genetics. In some aspects, the methods comprise: (a) extracting FeMV RNA from an isolated FeMV positive sample, (b) generating cDNAs from the FeMV RNA using primers that specifically hybridize to the FeMV RNA, (c) generating cDNA PCR amplicons from the cDNAs using primers that specifically hybridize to the cDNAs to produce cDNA PCR amplicons, (d) amplifying genomic and antigenomic termini of the FeMV RNA by rapid amplification of cDNA ends (RACE) using one or more RACE primers to produce RACE PCR amplicons, (e) purifying the cDNA PCR amplicons of step (c) and the RACE PCR amplicons of step ( d) to produce purified DNA, (f) sequencing the purified DNA to produce consensus sequences, (g) assembling the consensus sequences to produce a full-length FeMV genome, and (h) assembling the full-length FeMV genome in a plasmid.
Inventors:
DUPREX WILLIAM PAUL (US)
NAMBULLI SHAMKUMAR (US)
MURPHY LINDA J (US)
NAMBULLI SHAMKUMAR (US)
MURPHY LINDA J (US)
Application Number:
PCT/US2023/067540
Publication Date:
April 25, 2024
Filing Date:
May 26, 2023
Export Citation:
Assignee:
UNIV PITTSBURGH COMMONWEALTH SYS HIGHER EDUCATION (US)
International Classes:
C07K14/115; C12N5/10; C12N15/63; C12N15/86
Attorney, Agent or Firm:
SWANSON, Kristina L. et al. (US)
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