Strategy for efficient generation of numerous full-length cDNA clones of classical swine fever virus for haplotyping.

ABSTRACT:

Background

Direct molecular cloning of full-length cDNAs derived from viral RNA is an approach to identify the individual viral genomes within a virus population. This enables characterization of distinct viral haplotypes present during infection.

Results

In this study, we recover individual genomes of classical swine fever virus (CSFV), present in a pig infected with vKos that was rescued from a cDNA clone corresponding to the highly virulent CSFV Koslov strain. Full-length cDNA amplicons (ca. 12.3 kb) were made by long RT-PCR, using RNA extracted from serum, and inserted directly into a cloning vector prior to detailed characterization of the individual viral genome sequences. The amplicons used for cloning were deep sequenced, which revealed low level sequence variation (

SUBMITTER: Johnston CM 

PROVIDER: S-EPMC6085635 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature