| Autor: |
Lalruatfela, Bedekar, Megha Kadam, Godavarikar, Ankita, Valsalam, Anisha, Gireesh Babu, P., Rajendran, Kooloth Valappil |
| Předmět: |
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| Zdroj: |
Aquaculture International; Oct2024, Vol. 32 Issue 5, p5997-6015, 19p |
| Abstrakt: |
Tilapia lake virus (TiLV) is a novel single-stranded RNA virus that is considered a threat to the universal tilapia industry. Recombinant technology proved to apply to different viruses and does not require handling live viruses. The present study developed a recombinant protein from segment 4 of TiLV with the aim of developing an immunological detection method for the virus. For this purpose, the complete coding sequence of segment 4 of the virus was optimized, amplified, and cloned in pET-28a(+), a prokaryotic expression vector, and the TiLV segment 4 construct (pET-seg4) was developed. The recombinant protein was expressed in BL21-competent Escherichia coli by isopropyl-β-D-thiogalactopyranoside (IPTG) induction. The successful expression of the recombinant protein was confirmed by SDS-PAGE. The polyclonal antibody (PAb) raised against the recombinant protein was used for the indirect enzyme-linked immunosorbent assay (ELISA) development to detect TiLV in the pooled kidney and mucus samples. The coating concentrations for the recombinant protein and the TiLV-positive samples were standardized as 3.125 µg and 6.25 µg, respectively, and the optimum polyclonal antibody dilution was found as 1:800. The diagnostic sensitivity and diagnostic specificity of the assay were 82.35% and 100%, respectively. The recombinant protein developed in this study provided a better understanding of raising a codon-optimized protein and its use in developing an indirect ELISA test for TiLV, which can also be used as a recombinant vaccine for immunizing tilapia. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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