Accurate determination of the whole genome sequencing and open reading frames composition of non-replicating Tiantan strain of vaccinia virus based on novel long read sequencing platform
10.3760/cma.j.cn112309-20240201-00049
- VernacularTitle:基于新一代测序技术精准确定复制缺陷型痘苗病毒天坛株全基因组序列及其开放阅读框组成
- Author:
Zhongxian ZHANG
1
;
Changcheng WU
;
Han LI
;
Shuting HUO
;
Li ZHAO
;
Yao DENG
;
Xin MENG
;
Roujian LU
;
Houwen TIAN
;
Wenling WANG
;
Baoying HUANG
;
Wenjie TAN
Author Information
1. 包头医学院公共卫生学院，包头 014040
- Keywords:
Non-replicating Tiantan strain of vaccinia virus;
Third generation sequencing;
Whole genome sequencing;
Open reading frame
- From:
Chinese Journal of Microbiology and Immunology
2024;44(6):502-509
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To accurately ascertain the whole genome sequencing and the composition of open reading frames (ORFs) of non-replicating Tiantan strain of vaccinia virus (NTV) using next-generation long-read sequencing technology.Methods:NTV, obtained from our laboratory stock, was amplified and purified on chicken embryo fibroblast cells(CEFs), and the full-length genomic nucleic acid of NTV was extracted. The PacBio HiFi sequencing platform was utilized for de novo assembly to obtain the complete genomic sequence of NTV. Using a homology annotation strategy, we identified its ORF composition and compared it with known non-replicating vaccinia virus strains. Results:The total length of NTV′s genome was 171 729 bp, with a GC content of 33%. Its unique inverted terminal repeat (ITR) region comprised hairpin structures, two tandem repeat regions, and three non-repeat regions. NTV contained 166 ORFs, with major differences observed in the ITR and its surrounding regions when compared to MVA-BN and NYVAC. These three strains shared a common set of 138 ORFs. NTV encoded six unique ORFs related to virus evasion of host antiviral response.Conclusions:This study accurately determines the whole genome sequencing and ORFs composition of NTV, and reveals its similarities and differences with other replication-deficient vaccinia virus strains, which pave a way for the development and application of the next generation of monkeypox vaccines and novel viral vectors.
