THE DIFFERENTIAL GENE EXPRESSION OF LITOPENAEUS VANNAMEI AFTER WHITE SPOT SYNDROME VIRUS INFECTION BY DIFFERENTIAL DISPLAYED RT-PCR

Abstract

White leg shrimp (Litopenaeus vannamei) is the important economically aquatic animal of Thailand. White spot syndrome virus infection caused lost shrimp product and made an economic loss. This study aims to investigate the differential gene expression response to white spot syndrome virus infection in white leg shrimp. The quantity of white spot syndrome virus causes acute lethality within 3 days post-infection was an investigation in white leg shrimp. This dose was used to search for differential gene expression that response to white spot syndrome virus infection at the several timelines after white spot syndrome virus injection at 0, 6, 9, 12, 24 and 48 hour post-injection by differential displayed RT-PCR method by forty-five combinations between a two-bases anchored oligo-dT primer including AT, GT or CG and an arbitrary primer (P1 to P15).  The total 30 DNA bands which two times up-regulation from each pair of corresponding band was obtained from 9 combination primers.  The PCR product from the combination between two-base anchored oligo-dT primer with arbitrary primer obtained 1 band from oligo-dT-AT and P4, 17 bands from oligo-dT-CG and P1 P2 P3 and P4 whereas 12 bands from oligo-dT-GT. This selected-bands were then read for DNA sequence and match in GenBank database in the next study.