Evaluation of immunoprotective effects of recombinant proteins and DNA vaccines derived from Eimeria tenella surface antigen 6 and 15 in vivo

2021 ◽  
Author(s):  
Tiantian Geng ◽  
Liyan Luo ◽  
Yale Wang ◽  
Bang Shen ◽  
Rui Fang ◽  
...  
Keyword(s):  
Recombinant Proteins ◽  
Surface Antigen ◽  
Dna Vaccines ◽  
Eimeria Tenella

In vivo immunoprotective comparison between recombinant protein and DNA vaccine of Eimeria tenella surface antigen 4

2020 ◽  
Vol 278 ◽  
pp. 109032
Author(s):  
Pengfei Zhao ◽  
Yuncan Li ◽  
Yanqin Zhou ◽  
Junlong Zhao ◽  
Rui Fang
Keyword(s):  
Recombinant Protein ◽  
Dna Vaccine ◽  
Surface Antigen ◽  
Eimeria Tenella

scholarly journals DNA Vaccination by Electroporation and Boosting with Recombinant Proteins Enhances the Efficacy of DNA Vaccines for Schistosomiasis Japonica

2009 ◽  
Vol 16 (12) ◽  
pp. 1796-1803 ◽  
Author(s):  
Yang Dai ◽  
Yinchang Zhu ◽  
Donald A. Harn ◽  
Xiaoting Wang ◽  
Jianxia Tang ◽  
...  
Keyword(s):  
Adult Worm ◽  
Recombinant Proteins ◽  
Vaccine Efficacy ◽  
Dna Vaccines ◽  
Interleukin 2 ◽  
Schistosomiasis Japonica ◽  
Protein Vaccine ◽  
Control Programs ◽  
Protein Vaccines

ABSTRACT Schistosomiasis japonica is an endemic, zoonotic disease of major public health importance in China. Control programs combining chemotherapy and snail killing have not been able to block transmission of infection in lakes and marsh regions. Vaccination is needed as a complementary approach to the ongoing control programs. In the present study, we wanted to determine if the efficacies of DNA vaccines encoding the 23-kDa tetraspanin membrane protein (SjC23), triose phosphate isomerase (SjCTPI), and sixfold-repeated genes of the complementarity determining region 3 (CDR3) in the H chain of NP30 could be enhanced by boosting via electroporation in vivo and/or with cocktail protein vaccines. Mice vaccinated with cocktail DNA vaccines showed a significant worm reduction of 32.88% (P < 0.01) and egg reduction of 36.20% (P < 0.01). Vaccine efficacy was enhanced when animals were boosted with cocktail protein vaccines; adult worm and liver egg burdens were reduced 45.35% and 48.54%, respectively. Nearly identical results were obtained in mice boosted by electroporation in vivo, with adult worm and egg burdens reduced by 45.00% and 50.88%, respectively. The addition of a protein vaccine boost to this regimen further elevated efficacy to approximately 60% for adult worm burden and greater than 60% for liver egg reduction. The levels of interleukin-2, gamma interferon, and the ratios of immunoglobulin G2a (IgG2a)/IgG1 clearly showed that cocktail DNA vaccines induced CD4+ Th1-type responses. Boosting via either electroporation or with recombinant proteins significantly increased associated immune responses over those seen in mice vaccinated solely with DNA vaccines. Thus, schistosome DNA vaccine efficacy was significantly enhanced via boosting by electroporation in vivo and/or cocktail protein vaccines.

scholarly journals Covalent protein display on Hepatitis B core-like particles in plants through the in vivo use of the SpyTag/SpyCatcher system

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Hadrien Peyret ◽  
Daniel Ponndorf ◽  
Yulia Meshcheriakova ◽  
Jake Richardson ◽  
George P. Lomonossoff
Keyword(s):  
Hepatitis B ◽  
Cost Saving ◽  
Capsid Protein ◽  
Recombinant Proteins ◽  
Nicotiana Benthamiana ◽  
Vaccine Development ◽  
Binding Partner ◽  
Gfp Fluorescence ◽  
Display Systems

Abstract Virus-like particles (VLPs) can be used as nano-carriers and antigen-display systems in vaccine development and therapeutic applications. Conjugation of peptides or whole proteins to VLPs can be achieved using different methods such as the SpyTag/SpyCatcher system. Here we investigate the conjugation of tandem Hepatitis B core (tHBcAg) VLPs and the model antigen GFP in vivo in Nicotiana benthamiana. We show that tHBcAg VLPs could be successfully conjugated with GFP in the cytosol and ER without altering VLP formation or GFP fluorescence. Conjugation in the cytosol was more efficient when SpyCatcher was displayed on tHBcAg VLPs instead of being fused to GFP. This effect was even more obvious in the ER, showing that it is optimal to display SpyCatcher on the tHBcAg VLPs and SpyTag on the binding partner. To test transferability of the GFP results to other antigens, we successfully conjugated tHBcAg VLPs to the HIV capsid protein P24 in the cytosol. This work presents an efficient strategy which can lead to time and cost saving post-translational, covalent conjugation of recombinant proteins in plants.

scholarly journals Tat-SF1 Protein Associates with RAP30 and Human SPT5 Proteins

1999 ◽  
Vol 19 (9) ◽  
pp. 5960-5968 ◽  
Author(s):  
Jae B. Kim ◽  
Yuki Yamaguchi ◽  
Tadashi Wada ◽  
Hiroshi Handa ◽  
Phillip A. Sharp
Keyword(s):  
Recombinant Proteins ◽  
Transcriptional Activity ◽  
Transcription Activation ◽  
Cellular Proteins ◽  
Pol Ii ◽  
Nuclear Extracts ◽  
Cellular Factors ◽  
Immunodeficiency Virus

ABSTRACT The potent transactivator Tat recognizes the transactivation response RNA element (TAR) of human immunodeficiency virus type 1 and stimulates the processivity of elongation of RNA polymerase (Pol) II complexes. The cellular proteins Tat-SF1 and human SPT5 (hSPT5) are required for Tat activation as shown by immunodepletion with specific sera and complementation with recombinant proteins. In nuclear extracts, small fractions of both hSPT5 and Pol II are associated with Tat-SF1 protein. Surprisingly, the RAP30 protein of the heterodimeric transcription TFIIF factor is associated with Tat-SF1, while the RAP74 subunit of TFIIF is not coimmunoprecipitated with Tat-SF1. Overexpression of Tat-SF1 and hSPT5 specifically stimulates the transcriptional activity of Tat in vivo. These results suggest that Tat-SF1 and hSPT5 are indispensable cellular factors supporting Tat-specific transcription activation and that they may interact with RAP30 in controlling elongation.

scholarly journals Different expression levels of two KgmB-His fusion proteins

2005 ◽  
Vol 70 (12) ◽  
pp. 1401-1407 ◽  
Author(s):  
Sandra Markovic ◽  
Sandra Vojnovic ◽  
Milija Jovanovic ◽  
Branka Vasiljevic
Keyword(s):  
Recombinant Proteins ◽  
Fusion Proteins ◽  
Kanamycin Resistance ◽  
Expression Vectors ◽  
E Coli ◽  
C Terminus ◽  
N Terminus ◽  
Different Levels ◽  
Streptomyces Tenebrarius

The KgmB methylase from Streptomyces tenebrarius was expressed and purified using the QIAexpress System. Two expression vectors were made: pQEK-N, which places a (His)6 tag at the N-terminus, and pQEK-C, which places a (His)6 tag at the C-terminus of the recombinant KgmB protein. Kanamycin resistance of the E. coli cells containing either the pQEK-N or the pQEK-C recombinant plasmids confirmed the functionality of both KgmB-His fusion proteins in vivo. Interestingly, different levels of expression were observed between these two recombinant proteins. Namely, KgmB methylase with the (His)6 tag at the N-terminus showed a higher level of expression. Purification of the (His)6-tagged proteins using Ni-NTA affinity chromatography was performed under native conditions and the KgmB methylase with (His)6 tag at the N-terminus was purified to homogeneity >95 %. The recombinant KgmB protein was detected on a Western blot using anti-Sgm antibodies.

scholarly journals Detection of in vivo hepatitis B virus surface antigen mutations-A comparison of four routine screening assays

2018 ◽  
Vol 25 (10) ◽  
pp. 1132-1138 ◽  
Author(s):  
M. Gencay ◽  
A. Seffner ◽  
S. Pabinger ◽  
J. Gautier ◽  
P. Gohl ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Surface Antigen ◽  
Routine Screening ◽  
Virus Surface ◽  
Virus Surface Antigen ◽  
B Virus ◽  
Screening Assays

A Brief History of the Discovery of Amelogenin Nanoribbons In Vitro and In Vivo

2021 ◽  
pp. 002203452110434
Author(s):  
Y. Bai ◽  
J. Bonde ◽  
K.M.M. Carneiro ◽  
Y. Zhang ◽  
W. Li ◽  
...  
Keyword(s):  
Recombinant Proteins ◽  
Protein Structures ◽  
Historical Review ◽  
Limited Information ◽  
Transmission Electron Microscopy Analysis ◽  
Electron Microscopy Analysis ◽  
History Of ◽  
The 1960S

Without evidence for an organic framework, biological and biochemical processes observed during amelogenesis provided limited information on how extracellular matrix proteins control the development of the complex fibrous architecture of human enamel. Over a decade ago, amelogenin nanoribbons were first observed from recombinant proteins during in vitro mineralization experiments in our laboratory. In enamel from mice lacking the enzyme kallikrein 4 (KLK4), we later uncovered ribbon-like protein structures that matched the morphology, width, and thickness of the nanoribbons assembled by recombinant proteins. Interestingly, similar structures had already been described since the 1960s, when enamel sections from various mammals were demineralized and stained for transmission electron microscopy analysis. However, at that time, researchers were not aware of the ability of amelogenin to form nanoribbons and instead associated the filamentous nanostructures with possible imprints of mineral ribbons in the gel-like matrix of developing enamel. Further evidence for the significance of amelogenin nanoribbons for enamel development was stipulated when recent mineralization experiments succeeded in templating and orienting the growth of apatite ribbons along the protein nanoribbon framework. This article provides a brief historical review of the discovery of amelogenin nanoribbons in our laboratory in the context of reports by others on similar structures in the developing enamel matrix.

scholarly journals mRNA Engineering for the Efficient Chaperone-Mediated Co-Translational Folding of Recombinant Proteins in Escherichia coli

2019 ◽  
Vol 20 (13) ◽  
pp. 3163 ◽  
Author(s):  
Le Minh Bui ◽  
Almando Geraldi ◽  
Thi Thuy Nguyen ◽  
Jun Hyoung Lee ◽  
Ju Young Lee ◽  
...  
Keyword(s):  
Recombinant Proteins ◽  
Molecular Chaperones ◽  
Rna Binding ◽  
Protein Translation ◽  
Soluble Form ◽  
Mrna Transcript ◽  
Protein Substrates ◽  
Limited Success ◽  
Intergenic Sequences

The production of soluble, functional recombinant proteins by engineered bacterial hosts is challenging. Natural molecular chaperone systems have been used to solubilize various recombinant proteins with limited success. Here, we attempted to facilitate chaperone-mediated folding by directing the molecular chaperones to their protein substrates before the co-translational folding process completed. To achieve this, we either anchored the bacterial chaperone DnaJ to the 3ʹ untranslated region of a target mRNA by fusing with an RNA-binding domain in the chaperone-recruiting mRNA scaffold (CRAS) system, or coupled the expression of DnaJ and a target recombinant protein using the overlapping stop-start codons 5ʹ-TAATG-3ʹ between the two genes in a chaperone-substrate co-localized expression (CLEX) system. By engineering the untranslated and intergenic sequences of the mRNA transcript, bacterial molecular chaperones are spatially constrained to the location of protein translation, expressing selected aggregation-prone proteins in their functionally active, soluble form. Our mRNA engineering methods surpassed the in-vivo solubilization efficiency of the simple DnaJ chaperone co-overexpression method, thus providing more effective tools for producing soluble therapeutic proteins and enzymes.

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