scholarly journals Rescue of an Enterotropic Newcastle Disease Virus Strain ZM10 From Cloned cDNA and Stable Expressing an Inserted Foreign Gene

2021 ◽  
Author(s):  
Lei He ◽  
Hairong Wang ◽  
Zuhua Yu ◽  
Chengshui Liao ◽  
Ke Ding ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Reverse Genetics ◽  
Fluorescent Protein ◽  
Full Length ◽  
Foreign Gene ◽  
Ligation Independent Cloning ◽  
Recovery System ◽  
Cloned Cdna

Abstract Background: Newcastle disease virus (NDV) strain ZM10, a typical enterotropic avirulent vaccine strain, has been widely used in in China for chickens against Newcastle disease. To elucidate its enterotropic mechanism and develop recombiant multivalent vaccines based on it, the reverse genetics system for NDV ZM10 is an indispensable platform.Results: A full-length cDNA clone of NDV ZM10 and three supporting plasmids were constructed using the ligation-independent cloning (LIC) method. Recombinant NDV rZM10 was successfully rescued after these plasmids were co-transfected into BHK-21 cells. Besides, the recombinant virus rZM10-RFP encoding the red fluorescent protein was generated by inserting the RFP gene into the full-length clone of NDV between the P and M genes. These rescued viruses were genetically and biologically identical to the parental strain and showed similar growth kinetics. Conclusion: The recovery system of NDV ZM10 strain was established, and can be used as a foundation for research on the enterotropic mechanism and development of multivalent vaccines against viral diseases of livestock and poultry.

scholarly journals Expression of Two Foreign Genes from the Optimal Insertion Sites of Newcastle Disease Virus Vector for Use as a Multivalent Vaccine and Gene Therapy Vector

Proceedings ◽  
2020 ◽  
Vol 50 (1) ◽  
pp. 9
Author(s):  
Lei He ◽  
Zhenyu Zhang ◽  
Qingzhong Yu
Keyword(s):  
Gene Therapy ◽  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Fluorescent Protein ◽  
Transcription Unit ◽  
Lower Amount ◽  
Foreign Gene ◽  
Entry Site ◽  
Insertion Sites

Many Newcastle disease virus (NDV) strains have been developed as vectors to express a foreign gene (FG) for vaccine and gene therapy purposes. A majority of these NDV vectors express only a single FG or two FGs from suboptimal insertion sites in the NDV genome, obtaining various levels of FG expression. To improve the FG expression, we generated NDV LaSota vaccine strain-based recombinant viruses to express two FGs, green fluorescent protein (GFP) and red fluorescent protein (RFP) genes, from the identified optimal insertion sites, through a combination of the independent transcription unit (ITU) and the internal ribosomal entry site (IRES) dependent expression approaches. Biological assessments showed that these recombinants expressing two FGs were slightly attenuated with approximately one order of magnitude lower in virus titers than those containing a single FG. The FG expression efficiencies from two-FG viruses were also lower than those from the single-FG viruses. However, the expression of two FGs from the optimal insertion sites was significantly (p < 0.05) higher than those from the suboptimal insertion sites. The expression of FGs through the ITU approach was approximately 4-fold more efficient than that through the IRES-dependent approach. These results suggest that the NDV LaSota vector could efficiently express two FGs from the identified optimal insertions sites. The ITU strategy could be used for the expression of a higher amount of FG products, whereas the IRES tactic might be useful when a lower amount of FG products are needed.

scholarly journals High-level expression of a foreign gene from the most 3′-proximal locus of a recombinant Newcastle disease virus

2001 ◽  
Vol 82 (7) ◽  
pp. 1729-1736 ◽  
Author(s):  
Zhuhui Huang ◽  
Sateesh Krishnamurthy ◽  
Aruna Panda ◽  
Siba K. Samal
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Vaccine Vector ◽  
Foreign Gene ◽  
Gene Encoding ◽  
Reading Frame ◽  
Virulent Newcastle Disease Virus ◽  
Nucleocapsid Protein Gene ◽  
High Level

A previous report showed that insertion of a foreign gene encoding chloramphenicol acetyltransferase (CAT) between the HN and L genes of the full-length cDNA of a virulent Newcastle disease virus (NDV) yielded virus with growth retardation and attenuation. The NDV vector used in that study was pathogenic to chickens; it is therefore not suitable for use as a vaccine vector. In the present study, an avirulent NDV vector was generated and its potential to express CAT protein was evaluated. The CAT gene was under the control of NDV transcriptional start and stop signals and was inserted immediately before the open reading frame of the viral 3′-proximal nucleocapsid protein gene. A recombinant NDV expressing CAT activity at a high level was recovered. The replication and pathogenesis of the CAT-expressing recombinant NDV were not modified significantly. These results indicate the potential utility of an avirulent NDV as a vaccine vector.

Use of Reverse Genetics to Enhance the Oncolytic Properties of Newcastle Disease Virus

2007 ◽  
Vol 67 (17) ◽  
pp. 8285-8292 ◽  
Author(s):  
Adam Vigil ◽  
Man-Seong Park ◽  
Osvaldo Martinez ◽  
Mark A. Chua ◽  
Sa Xiao ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Reverse Genetics

scholarly journals Reverse Genetics Assembly of Newcastle Disease Virus Genome Template Using Asis-Sal-Pac BioBrick Strategy

2020 ◽  
Vol 22 (1) ◽  
Author(s):  
Amin Tavassoli ◽  
Safoura Soleymani ◽  
Alireza Haghparast ◽  
Gholamreza Hashemi Tabar ◽  
Mohammad Reza Bassami ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Reverse Genetics ◽  
Virus Genome

scholarly journals An improved method for the rescue of recombinant Newcastle disease virus

BioTechniques ◽  
2020 ◽  
Vol 68 (2) ◽  
pp. 96-100
Author(s):  
Pheik-Sheen Cheow ◽  
Tiong Kit Tan ◽  
Adelene Ai-Lian Song ◽  
Khatijah Yusoff ◽  
Suet Lin Chia
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Reverse Genetics ◽  
Vaccine Development ◽  
Transfection Efficiency ◽  
Transfection Method ◽  
Helper Plasmids ◽  
Immunogenic Properties ◽  
Recombinant Newcastle Disease Virus

Reverse genetics has been used to generate recombinant Newcastle disease virus with enhanced immunogenic properties for vaccine development. The system, which involves co-transfecting the viral antigenomic plasmid with three helper plasmids into a T7 RNA polymerase-expressing cell to produce viral progenies, poses a great challenge. We have modified the standard transfection method to improve the transfection efficiency of the plasmids, resulting in a higher titer of virus progeny production. Two transfection reagents (i.e., lipofectamine and polyethylenimine) were used to compare the transfection efficiency of the four plasmids. The virus progenies produced were quantitated with flow cytometry analysis of the infectious virus unit. The modified transfection method increased the titer of virus progenies compared with that of the standard transfection method.

Rescue of a recombinant Newcastle disease virus strain R2B expressing green fluorescent protein

Virus Genes ◽  
2017 ◽  
Vol 53 (3) ◽  
pp. 410-417 ◽  
Author(s):  
Madhan Mohan Chellappa ◽  
Sohini Dey ◽  
Satish Gaikwad ◽  
Dinesh C. Pathak ◽  
Vikram N. Vakharia
Keyword(s):  
Green Fluorescent Protein ◽  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Virus Strain ◽  
Newcastle Disease ◽  
Fluorescent Protein ◽  
Newcastle Disease Virus Strain ◽  
Green Fluorescent ◽  
Recombinant Newcastle Disease Virus
Sign in / Sign up

Export Citation Format

Share Document