Cloning, Expression, Purification, And Assessment Of Vibrio Parahaemolyticus-Agglutinating C-Type Lectin From Litopenaeus Vannamei Fused To GST Tag
Volume 4 - Issue 5, May 2020 Edition
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Phuong-Thao Thi Nguyen, Lan-Vy Hoai Nguyen, Quoc-Gia Mai, Thuoc Linh Tran, Hieu Tran-Van
AHPND, C-type lectin, Litopenaeus vannamei, LvLdlrCTL, recombinant protein, Vibrio parahaemolyticus
C-type lectins (CTLs) are a group of Pattern Recognition Receptors (PRR) that contains at least one carbohydrate recognition domain. CTLs play important roles in innate immunity of invertebrates by identifying and eliminating invading microorganisms. Recent studies have shown that a new type of CTL, LvLdrlCTL (Litopenaeus vannamei Low-Density Lipoprotein Receptor C-type Lectin) helps shrimp to resist to Acute Hepatopancreatic Necrosis Disease (AHPND) caused by Vibrio parahaemolyticus, which is currently no effective preventing and treatment available. LvLdlrCTL protein was fused with GST by cloning the LvLdlrCTL gene from Litopenaeus vannamei into pGEX-5X-1 vector and being expressed by host strain E. coli BL21 (DE3). The expression of GST-tagged LvLdlrCTL protein confirmed by SDS-PAGE and Western blot probed with GST antibody. The purified protein then was used for in vitro binding and agglutinating assay with V. paraheamolyticus. The result showed that GST-tagged LvLdlrCTL protein expressed in soluble form. Protein purification step could reach the purity above 85%. Lastly, purified protein could bind to and agglutinate AHPND-causing V. parahaemolyticus. This initial trial created the groundwork for expression and purification in large quantities of LvLdlrCTL. The recombinant protein could be used for further developing on anti-AHPND in white leg shrimps.
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