Cargando…

Construction of an easily detectable transgenic Synechococcus elongatus PCC 7942 against White Spot Syndrome Virus using vp28 and mOrange Gene and its metabolism in shrimp

White spot syndrome is an epidemic disease caused by the highly contagious and lethal white spot syndrome virus (WSSV), resulting in huge economic losses to the global aquaculture industry. VP28 is the main structural protein in the capsule of WSSV and is important in the early stage of infection. U...

Descripción completa

Detalles Bibliográficos
Autores principales: Peng, Wen, Bao, Qunjing, Jia, Rui, He, Peimin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9459150/
https://www.ncbi.nlm.nih.gov/pubmed/36091009
http://dx.doi.org/10.3389/fimmu.2022.974014
_version_ 1784786442081271808
author Peng, Wen
Bao, Qunjing
Jia, Rui
He, Peimin
author_facet Peng, Wen
Bao, Qunjing
Jia, Rui
He, Peimin
author_sort Peng, Wen
collection PubMed
description White spot syndrome is an epidemic disease caused by the highly contagious and lethal white spot syndrome virus (WSSV), resulting in huge economic losses to the global aquaculture industry. VP28 is the main structural protein in the capsule of WSSV and is important in the early stage of infection. Under an excitation wavelength of 548 nm, the mOrange fluorescent protein releases a 562 nm emission wavelength, which is different from the autofluorescence of cyanobacteria. Therefore, using this characteristic combined with the receptor system of Synechococcus elongatus PCC 7942, we constructed transgenic S. elongatus to express the recombinant protein VP28-mOrange. In addition, PCR and western blotting were used to confirm the stable expression of the target gene in cyanobacteria. Using mOrange tracer features, we explored the recombinant protein VP28-mOrange in the metabolic cycle of young Litopenaeus Vannamei after feeding. After the young shrimp had stopped consuming transgenic cyanobacteria, the 24 to 33 h fluorescence signal in the intestine was very weak, and almost disappeared after 36 h. We explored the protective effect of transgenic vp28-mOrange S. elongatus within 48 h of being ingested by L. vannamei and set WSSV challenges at 2, 12, 24, and 48 h post-immunization. However, the survival rate of L. vannamei decreased as the time of the WSSV challenge increased. The survival rate on the seventh day was 81%, 52%, 45.5%, and 33.3% for shrimps challenged for 2, 12, 24, and 48 h, respectively. Enzyme activity can also support this conjecture, the enzyme activity indexes of the experimental groups were significantly reduced compared to positive and wild-type controls. Therefore, this immune agent functioned as a preventive agent. Compared with the traditional method, this method was easy to detect and can visualize the digestion of transgenic cyanobacteria in the Litopenaeus vannamei intestine.
format Online
Article
Text
id pubmed-9459150
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-94591502022-09-10 Construction of an easily detectable transgenic Synechococcus elongatus PCC 7942 against White Spot Syndrome Virus using vp28 and mOrange Gene and its metabolism in shrimp Peng, Wen Bao, Qunjing Jia, Rui He, Peimin Front Immunol Immunology White spot syndrome is an epidemic disease caused by the highly contagious and lethal white spot syndrome virus (WSSV), resulting in huge economic losses to the global aquaculture industry. VP28 is the main structural protein in the capsule of WSSV and is important in the early stage of infection. Under an excitation wavelength of 548 nm, the mOrange fluorescent protein releases a 562 nm emission wavelength, which is different from the autofluorescence of cyanobacteria. Therefore, using this characteristic combined with the receptor system of Synechococcus elongatus PCC 7942, we constructed transgenic S. elongatus to express the recombinant protein VP28-mOrange. In addition, PCR and western blotting were used to confirm the stable expression of the target gene in cyanobacteria. Using mOrange tracer features, we explored the recombinant protein VP28-mOrange in the metabolic cycle of young Litopenaeus Vannamei after feeding. After the young shrimp had stopped consuming transgenic cyanobacteria, the 24 to 33 h fluorescence signal in the intestine was very weak, and almost disappeared after 36 h. We explored the protective effect of transgenic vp28-mOrange S. elongatus within 48 h of being ingested by L. vannamei and set WSSV challenges at 2, 12, 24, and 48 h post-immunization. However, the survival rate of L. vannamei decreased as the time of the WSSV challenge increased. The survival rate on the seventh day was 81%, 52%, 45.5%, and 33.3% for shrimps challenged for 2, 12, 24, and 48 h, respectively. Enzyme activity can also support this conjecture, the enzyme activity indexes of the experimental groups were significantly reduced compared to positive and wild-type controls. Therefore, this immune agent functioned as a preventive agent. Compared with the traditional method, this method was easy to detect and can visualize the digestion of transgenic cyanobacteria in the Litopenaeus vannamei intestine. Frontiers Media S.A. 2022-08-26 /pmc/articles/PMC9459150/ /pubmed/36091009 http://dx.doi.org/10.3389/fimmu.2022.974014 Text en Copyright © 2022 Peng, Bao, Jia and He https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Peng, Wen
Bao, Qunjing
Jia, Rui
He, Peimin
Construction of an easily detectable transgenic Synechococcus elongatus PCC 7942 against White Spot Syndrome Virus using vp28 and mOrange Gene and its metabolism in shrimp
title Construction of an easily detectable transgenic Synechococcus elongatus PCC 7942 against White Spot Syndrome Virus using vp28 and mOrange Gene and its metabolism in shrimp
title_full Construction of an easily detectable transgenic Synechococcus elongatus PCC 7942 against White Spot Syndrome Virus using vp28 and mOrange Gene and its metabolism in shrimp
title_fullStr Construction of an easily detectable transgenic Synechococcus elongatus PCC 7942 against White Spot Syndrome Virus using vp28 and mOrange Gene and its metabolism in shrimp
title_full_unstemmed Construction of an easily detectable transgenic Synechococcus elongatus PCC 7942 against White Spot Syndrome Virus using vp28 and mOrange Gene and its metabolism in shrimp
title_short Construction of an easily detectable transgenic Synechococcus elongatus PCC 7942 against White Spot Syndrome Virus using vp28 and mOrange Gene and its metabolism in shrimp
title_sort construction of an easily detectable transgenic synechococcus elongatus pcc 7942 against white spot syndrome virus using vp28 and morange gene and its metabolism in shrimp
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9459150/
https://www.ncbi.nlm.nih.gov/pubmed/36091009
http://dx.doi.org/10.3389/fimmu.2022.974014
work_keys_str_mv AT pengwen constructionofaneasilydetectabletransgenicsynechococcuselongatuspcc7942againstwhitespotsyndromevirususingvp28andmorangegeneanditsmetabolisminshrimp
AT baoqunjing constructionofaneasilydetectabletransgenicsynechococcuselongatuspcc7942againstwhitespotsyndromevirususingvp28andmorangegeneanditsmetabolisminshrimp
AT jiarui constructionofaneasilydetectabletransgenicsynechococcuselongatuspcc7942againstwhitespotsyndromevirususingvp28andmorangegeneanditsmetabolisminshrimp
AT hepeimin constructionofaneasilydetectabletransgenicsynechococcuselongatuspcc7942againstwhitespotsyndromevirususingvp28andmorangegeneanditsmetabolisminshrimp