scholarly journals Recovery of Recombinant Avian Paramyxovirus Type-3 Strain Wisconsin by Reverse Genetics and Its Evaluation as a Vaccine Vector for Chickens

Viruses ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 316
Author(s):  
Mohamed A. Elbehairy ◽  
Sunil K. Khattar ◽  
Siba K. Samal
Keyword(s):  
Growth Rate ◽  
Tissue Distribution ◽  
Vaccine Vector ◽  
Expression Level ◽  
Reverse Genetic System ◽  
Foreign Protein ◽  
Foreign Gene ◽  
Avian Paramyxovirus ◽  
Gfp Expression ◽  
Type 3

A reverse genetic system for avian paramyxovirus type-3 (APMV-3) strain Wisconsin was created and the infectious virus was recovered from a plasmid-based viral antigenomic cDNA. Green fluorescent protein (GFP) gene was cloned into the recombinant APMV-3 genome as a foreign gene. Stable expression of GFP by the recovered virus was confirmed for at least 10 consecutive passages. APMV-3 strain Wisconsin was evaluated against APMV-3 strain Netherlands and APMV-1 strain LaSota as a vaccine vector. The three viral vectors expressing GFP as a foreign protein were compared for level of GFP expression level, growth rate in chicken embryo fibroblast (DF-1) cells, and tissue distribution and immunogenicity in specific pathogen-free (SPF) day-old chickens. APMV-3 strain Netherlands showed highest growth rate and GFP expression level among the three APMV vectors in vitro. APMV-3 strain Wisconsin and APMV-1 strain LaSota vectors were mainly confined to the trachea after vaccination of day-old SPF chickens without any observable pathogenicity, whereas APMV-3 strain Netherlands showed wide tissue distribution in different body organs (brain, lungs, trachea, and spleen) with mild observable pathogenicity. In terms of immunogenicity, both APMV-3 strain-vaccinated groups showed HI titers two to three fold higher than that induced by APMV-1 strain LaSota vaccinated group. This study offers a novel paramyxovirus vector (APMV-3 strain Wisconsin) which can be used safely for vaccination of young chickens as an alternative for APMV-1 strain LaSota vector.

Systematic analysis of attenuated Coxsackievirus expressing a foreign gene as a viral vaccine vector

Vaccine ◽  
2010 ◽  
Vol 28 (5) ◽  
pp. 1234-1240 ◽  
Author(s):  
Dae-Sun Kim ◽  
Young-Joo Cho ◽  
Byoung-Guk Kim ◽  
Su-Hae Lee ◽  
Jae-Hwan Nam
Keyword(s):  
Vaccine Vector ◽  
Foreign Gene ◽  
Systematic Analysis ◽  
Viral Vaccine

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.

scholarly journals Expression Pattern of the Alpha-Kafirin Promoter Coupled with a Signal Peptide fromSorghum bicolorL. Moench

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Norazlina Ahmad ◽  
Rajnesh Sant ◽  
Milovan Bokan ◽  
Kathryn J. Steadman ◽  
Ian D. Godwin
Keyword(s):  
Gene Expression ◽  
Signal Peptide ◽  
Sweet Corn ◽  
Signal Sequence ◽  
Seed Storage ◽  
Foreign Gene ◽  
Regulatory Sequences ◽  
Fluorescent Properties ◽  
Gfp Expression ◽  
Foreign Gene Expression

Regulatory sequences with endosperm specificity are essential for foreign gene expression in the desired tissue for both grain quality improvement and molecular pharming. In this study, promoters of seed storage α-kafirin genes coupled with signal sequence (ss) were isolated fromSorghum bicolorL. Moench genomic DNA by PCR. The α-kafirin promoter (α-kaf) contains endosperm specificity-determining motifs, prolamin-box, the O2-box 1, CATC, and TATA boxes required for α-kafirin gene expression in sorghum seeds. The constructs pMB-Ubi-gfpand pMB-kaf-gfpwere microprojectile bombarded into various sorghum and sweet corn explants. GFP expression was detected on all explants using the Ubi promoter but only in seeds for the α-kaf promoter. This shows that the α-kaf promoter isolated was functional and demonstrated seed-specific GFP expression. The constructs pMB-Ubi-ss-gfpand pMB-kaf-ss-gfpwere also bombarded into the same explants. Detection of GFP expression showed that the signal peptide (SP)::GFP fusion can assemble and fold properly, preserving the fluorescent properties of GFP.

scholarly journals Consequences of prenatal and preweaning growth for yield of beef primal cuts from 30-month-old Piedmontese- and Wagyu-sired cattle

2009 ◽  
Vol 49 (6) ◽  
pp. 468 ◽  
Author(s):  
P. L. Greenwood ◽  
L. M. Cafe ◽  
H. Hearnshaw ◽  
D. W. Hennessy ◽  
S. G. Morris
Keyword(s):  
Growth Rate ◽  
Environmental Factors ◽  
Tissue Distribution ◽  
Early Life ◽  
Low Birthweight ◽  
Carcass Weight ◽  
Whole Body ◽  
Target Market ◽  
Subsequent Growth

Cattle sired by Piedmontese or Wagyu bulls were bred and grown within pasture-based nutritional systems followed by feedlot finishing. Effects of low (mean 28.6 kg, n = 120) and high (38.8 kg, n = 120) birthweight followed by slow (mean 554 g/day, n = 119) or rapid (875 g/day, n = 121) growth to weaning on beef primal cut weights at ~30 months of age were examined. Cattle of low birthweight or grown slowly to weaning had smaller primal cuts at 30 months as a result of reduced liveweight and smaller carcasses compared with their high birthweight or rapidly grown counterparts. Hence they require additional nutritional and economic inputs to reach target market weights. At equivalent carcass weights (380 kg), cattle restricted in growth from birth to weaning yielded slightly more beef and were somewhat leaner than their rapidly grown counterparts, resulting in primal cuts being up to 4% heavier in the slowly grown compared with the rapidly grown cattle. Compositional differences due to birthweight were less apparent at the same carcass weight, although low birthweight cattle had a slightly (~2%) heavier forequarter and slightly lower (~1%) hindquarter retail yield, and less shin-shank meat (~2%) than high birthweight cattle, suggesting only minor effects on carcass tissue distribution. There were few interactions between sire genotype and birthweight or preweaning growth, and interactions between birthweight and preweaning growth were not evident for any variables. However, variability between cohorts in their long-term responses to growth early in life suggests other environmental factors during early-life and/or subsequent growth influenced carcass yield characteristics. Overall, this study shows that effects of birthweight and preweaning growth rate on carcass compositional and yield characteristics were mostly explained by variation in carcass weight and, hence, in whole body growth to 30 months of age.

Turkey avian paramyxovirus type 3 infections in poultry

1990 ◽  
Vol 46 (3) ◽  
pp. 211-218 ◽  
Author(s):  
I.P.R. Awang ◽  
P.H. Russell
Keyword(s):  
Avian Paramyxovirus ◽  
Type 3

scholarly journals Regulating the T7 RNA polymerase expression in E. coli BL21 (DE3) to provide more host options for recombinant protein production

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Fei Du ◽  
Yun-Qi Liu ◽  
Ying-Shuang Xu ◽  
Zi-Jia Li ◽  
Yu-Zhou Wang ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Rna Polymerase ◽  
Recombinant Proteins ◽  
Expression System ◽  
T7 Rna Polymerase ◽  
Expression Level ◽  
Foreign Protein ◽  
Atpase Subunit ◽  
E Coli ◽  
Prokaryotic Expression System

AbstractEscherichia coli is the most widely used bacterium in prokaryotic expression system for the production of recombinant proteins. In BL21 (DE3), the gene encoding the T7 RNA polymerase (T7 RNAP) is under control of the strong lacUV5 promoter (PlacUV5), which is leakier and more active than wild-type lac promoter (PlacWT) under certain growth conditions. These characteristics are not advantageous for the production of those recombinant proteins with toxic or growth-burdened. On the one hand, leakage expression of T7 RNAP leads to rapid production of target proteins under non-inducing period, which sucks resources away from cellular growth. Moreover, in non-inducing or inducing period, high expression of T7 RNAP production leads to the high-production of hard-to-express proteins, which may all lead to loss of the expression plasmid or the occurrence of mutations in the expressed gene. Therefore, more BL21 (DE3)-derived variant strains with rigorous expression and different expression level of T7 RNAP should be developed. Hence, we replaced PlacUV5 with other inducible promoters respectively, including arabinose promoter (ParaBAD), rhamnose promoter (PrhaBAD), tetracycline promoter (Ptet), in order to optimize the production of recombinant protein by regulating the transcription level and the leakage level of T7 RNAP. Compared with BL21 (DE3), the constructed engineered strains had higher sensitivity to inducers, among which rhamnose and tetracycline promoters had the lowest leakage ability. In the production of glucose dehydrogenase (GDH), a protein that causes host autolysis, the engineered strain BL21 (DE3::ara) exhibited higher biomass, cell survival rate and foreign protein expression level than that of BL21 (DE3). In addition, these engineered strains had been successfully applied to improve the production of membrane proteins, including E. coli cytosine transporter protein (CodB), the E. coli membrane protein insertase/foldase (YidC), and the E. coli F-ATPase subunit b (Ecb). The engineered strains constructed in this paper provided more host choices for the production of recombinant proteins.

scholarly journals Transfer of Green Fluorescent Protein (GFP) Gene to Betta splendens Embryos by Transfection and Electroporation Methods

Omni-Akuatika ◽  
2018 ◽  
Vol 14 (2) ◽  
Author(s):  
Eni Kusrini ◽  
Alimuddin Alimuddin ◽  
Erma Primanita Hayuningtyas ◽  
Syuhada Restu Danupratama
Keyword(s):  
Green Fluorescent Protein ◽  
Gene Transfer ◽  
Expression Vector ◽  
Fluorescent Protein ◽  
Betta Splendens ◽  
Foreign Gene ◽  
Gfp Expression ◽  
Dna Concentration ◽  
Gfp Gene ◽  
Green Fluorescent

Transfection and electroporation method shave a high possibility to apply towards transgenic production of small eggs size fish species.  This study aimed to examine the potential of transfection and electroporation methods to use for transferring a foreign gene into betta fish (Betta splendens) embryos using green fluorescent protein (GFP) gene as a model.  Fish were spawned naturally in the ratio of male: female was 1:1, then a total of 200 eggs were taken for each treatment.  Transfection was performed for 30 minutes (room temperature of about 25 °C) at two-cell stage of embryos using transfast reagent.  Transfection reaction consisted of 0.75 µL transfast reagent, 0.25 µL GFP expression vector (DNA concentration: 50 µg/µL) and 99 µL NaCl solution (concentration: 0,95%).  Electroporation was performed using 125 volt cm-1, 3 times pulse frequency at one second interval and pulse length of 7 micro seconds.  A volume of 800 µL GFP expression vector solution (DNA concentration: 50 µg/ µL) in PBS was used for electroporation.  The successful of foreign gene transfer was determined by PCR method with GFP specific primers.  The results showed that hatching rate of eggs in transfection treatment was 67.08%, while the electroporation was 72.09%.  Survival of larvae in transfection treatment was 73.00%, while the electroporation was 75.00%.  The results of PCR analysis showed that transfection method allowed 65% of the survived fish carrying GFP gene, whereas the electroporation method was 70%.  Thus, foreign gene transfer in betta fish can be conducted using the transfection and electroporation methods. 

scholarly journals Development of a Novel Avian Vaccine Vector Derived From the Emerging Fowl Adenovirus 4

2021 ◽  
Vol 12 ◽  
Author(s):  
Qing Pan ◽  
Yu Zhang ◽  
Aijing Liu ◽  
Hongyu Cui ◽  
Yulong Gao ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Virus Genome ◽  
Open Reading Frames ◽  
Vaccine Vector ◽  
Economic Losses ◽  
Foreign Gene ◽  
The Novel ◽  
Hydropericardium Syndrome ◽  
Fowl Adenovirus ◽  
Green Fluorescent

Severe hepatitis-hydropericardium syndrome (HHS) associated with a novel viral genotype, fowl adenovirus 4 (FAdV-4), has emerged and widely spread in China since 2015, causing severe economic losses to the poultry industry. We previously reported that the hexon gene is responsible for pathogenicity and obtained a non-pathogenic hexon-replacement rHN20 strain; however, the lack of information about the non-essential regions for virus replication limits the development of a FAdV-4 vector. This study first established an enhanced green fluorescent protein (EGFP)-indicator virus based on the FAdV-4 reverse genetic technique, effective for batch operations in the virus genome. Based on this, 10 open reading frames (ORFs) at the left end and 13 ORFs at the right end of the novel FAdV-4 genome were deleted separately and identified as non-essential genes for viral replication, providing preliminary insertion sites for foreign genes. To further improve its feasibility as a vaccine vector, seven combinations of ORFs were successfully replaced with EGFP without affecting the immunogenicity of the vector backbone. Finally, a recombinant rHN20-vvIBDV-VP2 strain, expressing the VP2 protein of very virulent infectious bursa disease virus (vvIBDV), was rescued and showed complete protection against FAdV-4 and vvIBDV. Thus, the novel FAdV-4 vector could provide sufficient protection for HHS and efficient exogenous gene delivery. Overall, our findings systemically identified 23 non-essential ORFs for FAdV-4 replication and seven foreign gene insertion regions, providing valuable information for an in-depth understanding of the novel FAdV-4 pathogenesis and development of multivalent vaccines.

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