Characterization of an Australian isolate of taro bacilliform virus and development of an infectious clone

AbstractAn unprecedented number of viruses have been discovered by leveraging advances in high-throughput sequencing. Infectious clone technology is a universal approach that facilitates the study of biology and role in disease of viruses. In recent years homology-based cloning methods such as Gibson assembly have been used to generate virus infectious clones. We detail herein the preparation of home-made cloning materials for Gibson assembly. The home-made materials were used in one-step generation of the infectious cDNA clone of a plant RNA virus into a T-DNA binary vector. The clone was verified by a single Illumina reaction and a de novo read assembly approach that required no primer walking, custom primers or reference sequences. Clone infectivity was finally confirmed by Agrobacterium-mediated delivery to host plants. We anticipate that the convenient home-made materials, one-step cloning and Illumina verification strategies described herein will accelerate characterization of viruses and their role in disease development.

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Construction and characterization of a full-length cDNA infectious clone of emerging porcine Senecavirus A

Virology ◽

10.1016/j.virol.2016.07.003 ◽

2016 ◽

Vol 497 ◽

pp. 111-124 ◽

Cited By ~ 22

Author(s):

Zhenhai Chen ◽

Fangfeng Yuan ◽

Yanhua Li ◽

Pengcheng Shang ◽

Robin Schroeder ◽

...

Keyword(s):

Infectious Clone ◽

Full Length ◽

Full Length Cdna

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Construction and characterization of the infectious clone of porcine parvovirus carrying genetic marker

Veterinary Microbiology ◽

10.1016/j.vetmic.2019.06.015 ◽

2019 ◽

Vol 235 ◽

pp. 143-150 ◽

Cited By ~ 1

Author(s):

Songbiao Chen ◽

Bichen Miao ◽

Huan Zhang ◽

Yingli Xiong ◽

Xiujuan Zhang ◽

...

Keyword(s):

Genetic Marker ◽

Infectious Clone ◽

Porcine Parvovirus

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Identification of Canine Coronavirus Strains from Feces by S Gene Nested PCR and Molecular Characterization of a New Australian Isolate

Journal of Clinical Microbiology ◽

10.1128/jcm.39.3.1036-1041.2001 ◽

2001 ◽

Vol 39 (3) ◽

pp. 1036-1041 ◽

Cited By ~ 21

Author(s):

M. J. Naylor ◽

G. A. Harrison ◽

R. P. Monckton ◽

S. McOrist ◽

P. R. Lehrbach ◽

...

Keyword(s):

Molecular Characterization ◽

Nested Pcr ◽

Australian Isolate ◽

S Gene ◽

Canine Coronavirus

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Construction and characterization of a full-length infectious clone from a fast-replicating, X4-tropic HIV-1 subtype B′ isolate

Archives of Virology ◽

10.1007/s00705-010-0776-z ◽

2010 ◽

Vol 155 (12) ◽

pp. 1923-1931 ◽

Cited By ~ 2

Author(s):

Shou-li Wu ◽

Yan-sheng Yan ◽

Ping-ping Yan ◽

Hai-long Huang ◽

Hui-rong Wang

Keyword(s):

Infectious Clone ◽

Full Length ◽

Subtype B ◽

Hiv 1

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Development and characterization of a clinical strain of Coxsackievirus A16 and an eGFP infectious clone

Virologica Sinica ◽

10.1007/s12250-015-3610-7 ◽

2015 ◽

Vol 30 (4) ◽

pp. 269-276 ◽

Cited By ~ 4

Author(s):

Chenglin Deng ◽

Xiaodan Li ◽

Siqing Liu ◽

Linlin Xu ◽

Hanqing Ye ◽

...

Keyword(s):

Infectious Clone ◽

Clinical Strain ◽

Coxsackievirus A16

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Development and characterization of a double subgenomic chikungunya virus infectious clone to express heterologous genes in Aedes aegypti mosqutioes

Insect Biochemistry and Molecular Biology ◽

10.1016/j.ibmb.2005.05.008 ◽

2005 ◽

Vol 35 (10) ◽

pp. 1162-1170 ◽

Cited By ~ 56

Author(s):

Dana L. Vanlandingham ◽

Konstantin Tsetsarkin ◽

Chao Hong ◽

Kimberly Klingler ◽

Kate L. McElroy ◽

...

Keyword(s):

Aedes Aegypti ◽

Chikungunya Virus ◽

Infectious Clone

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Development and characterization of infectious clones of two strains of Usutu virus

10.1101/2020.08.05.238543 ◽

2020 ◽

Author(s):

Tyler Bates ◽

Christina Chuong ◽

Seth A. Hawks ◽

Pallavi Rai ◽

Rebecca M. Salgado ◽

...

Keyword(s):

Reverse Genetics ◽

Insect Cells ◽

Rna Virus ◽

Infectious Clone ◽

Bird Species ◽

Infection Transmission ◽

Usutu Virus ◽

Infectious Clones ◽

Positive Sense

AbstractUsutu virus (USUV; genus Flavivirus; family Flaviviridae) is a mosquito-borne, positive-sense RNA virus that is currently causing significant die-offs in numerous bird species throughout Europe and has caused infections in humans. Currently, there are no molecular clones for USUV, hence, hindering studies on the pathogenesis and transmission of USUV. In this report, we demonstrate the development and characterization of infectious clones for two modern strains of USUV isolated from Europe and Africa. We show that the infectious clone-derived viruses replicated similarly to the parental strains in both mammalian and insect cells. Additionally, we observed similar levels of replication and pathogenesis in two mouse models. This reverse genetics system will aid the scientific community in studying and developing USUV infection, transmission, diagnostics, and vaccines.

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