Deeply Sequenced Infectious Clones of Key Cassava Begomovirus Isolates from Cameroon.

J Steen Hoyer,Vincent N Fondong,David O Deppong,Catherine Doyle Aimone,Linda Hanley-Bowdoin,Mary M Dallas,Siobain Duffy,Kenneth M Stedman

doi:10.1128/mra.00802-20

Abstract

We deeply sequenced two pairs of widely used infectious clones (4 plasmids) of the bipartite begomoviruses African cassava mosaic virus (ACMV) and East African cassava mosaic Cameroon virus (EACMCV). The ACMV clones were quite divergent from published sequences. Raw reads, consensus plasmid sequences, and the infectious clones themselves are all publicly available.

Highlights

Infectious clones are a central tool of molecular virology
We describe new sequence resources for these clones, which essentially confirm the sequence of the East African cassava mosaic Cameroon virus (EACMCV) clones and clarify the identity of the African cassava mosaic virus (ACMV) clones
Data are available as SRA BioProject PRJNA649777. Three of these plasmids were described by Fondong et al (2000), and the fourth, for EACMCV DNA-B, was described later (Chowda Reddy et al, 2012; Fondong and Chen, 2011) due to greater difficulty in cloning that sequence in the partial-tandem-dimer configuration

Summary

Introduction

Circular single-stranded DNA viruses such as begomoviruses are often cloned in a two-step process to create partial tandem dimers containing two copies of the virus origin-of-replication hairpin. These clones have been used extensively for molecular genetic analysis (Amin et al, 2011; Beyene et al, 2016; Chauhan et al, 2018; Chellappan et al, 2004, 2005a, 2005b; Chowda Reddy et al, 2008, 2009, 2012; Fondong et al, 2007; Kuria et al, 2017; Ndunguru et al, 2016; Patil et al, 2016; Pita et al, 2001; Reyes et al, 2013; Vanitharani et al, 2003, 2004), including analysis of spontaneous mutation (Aimone et al, 2020; Chen et al, 2019; Fondong and Chen, 2011).

Results

Conclusion