Determination of transgene stability in Nannochloropsis sp. transformed with immunogenic peptide for oral vaccination against vibriosis

Aisamuddin Ardi Zainal Abidin,Zetty Norhana Balia Yusof,Fatimah Md Yusoff,Nur Adila Othman

doi:10.1007/s10499-020-00634-w

Aisamuddin Ardi Zainal Abidin, Zetty Norhana Balia Yusof + Show 2 more

Open Access

https://doi.org/10.1007/s10499-020-00634-w

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Journal: Aquaculture International	Publication Date: Jan 20, 2021
Citations: 7	License type: open-access

Affiliation: Universiti Putra Malaysia

Abstract

Vibriosis is one of the common diseases caused by gram-negative bacteria from the genus Vibrio. To treat vibriosis, vaccination has been proven to be the most effective treatment as it can avoid the risk of drugs or antibiotics resistance. Microalgae are commonly used as feed for aquatic organisms and Nannochloropsis sp. is one of the highly utilized species for fish feed. This study focused on the use of microalga, Nannochloropsis sp. as a vaccine carrier. Transgenic Nannochloropsis sp. harbouring an outer membrane protein kinase (OmpK) gene fragment of the Vibrio species namely V1, V2, CV1, CV2, CPV1 and CPV2 were utilized in this study. The stability of OmpK gene in transgenic Nannochloropsis sp. over a number of generations was evaluated. DNA and RNA from the Nannochloropsis sp. transgenic lines were extracted and subjected to PCR amplification of OmpK gene fragment. The OmpK gene fragment was successfully amplified and expressed up to the fifth generation (F5). For V1, V2, CV1 and CV2, the gene was present and expressed in fourth generation (F4) and F5 respectively but CPV1 and CPV2 the OmpK genes were present up to F4. From the results obtained, Nannochloropsis sp. is shown to be suitable as a vaccine carrier and can be utilized as a vaccine carrier to ameliorate vibriosis.

Highlights

Aquaculture is one of the industries that contribute to world’s economy where the global market for aquaculture products is about $50 billion over a year (Chen et al 2008)
DNAs extracted from the transgenic lines were analysed using polymerase chain reaction (PCR) to amplify the homologous region where hypothetically is the site of integration
Analysis using PCR showed that survived transgenic lines of Nannochloropsis sp. transformants contain the expression cassette which is indicated by the single band between 10 and 1.5 kb (Fig. 4)

Summary

Introduction

Aquaculture is one of the industries that contribute to world’s economy where the global market for aquaculture products is about $50 billion over a year (Chen et al 2008). Disease is the major problem in the aquaculture industry where it causes high economic losses for the industry. Bacterial infection in various aquatic animal constitutes the most important source of disease problems while fungal is the second major cause of losses in aquaculture (Meyer 1991). To control vibriosis in aquaculture, antibiotics are widely used in various ways such as incorporation into feed as well as oral administration (Shoemaker et al 2009). These approaches can cause other problems in aquatic animals and in humans and the environment. Different strategies need to be developed in order to handle this situation

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