Profiling of differentially expressed genes in hepatopancreas of white spot syndrome virus-resistant shrimp (Litopenaeus vannamei) by suppression subtractive hybridisation.

摘要

In order to find immune-relevant factors responsible for virus resistance and response to the virus infection, the suppression subtractive hybridisation method was employed to identify differentially expressed genes and their expression profiles in the hepatopancreas of the white spot syndrome virus (WSSV) resistant and susceptible Pacific white shrimp (Litopenaeus vannamei). Two forward subtractive libraries (at 0 and 48 h time point) and two reverse subtractive libraries (at 0 and 48 h time point) were constructed, and more than 1200 clones were sequenced, of which 40 differentially expressed genes were identified. These genes encode proteins corresponding to a wide range of functions, including defence-related proteins, enzymes, transcription factors, apoptotic-related proteins, intracellular components potentially related to signaling cascades, metabolic proteins, and cytoskeletal protein. Five genes (laccase, carboxypeptidase B, H+-transporting ATP synthase, Acyl-ConA-binding protein (ACBP), and cortical granule protein with LDL-receptor) are found for the first time in shrimp and their expressions were up-regulated in the virus-resistant shrimp. Among the 40 genes, 30 showed up-regulation in the virus-resistant shrimp comparing with susceptible shrimp, while 10 genes showed down-regulation. Haemocyanin was the most abundant gene in our forward subtractive libraries. In addition, chathepsin L, ecdysteroid regulated protein, zinc proteinase, lectin, sterol carrier protein-X, lysozyme, cortical granule protein with LDL-receptor, leucine-rich repeat LGI family, fatty acid binding protein, and preamylase all showed up-regulation in the resistant shrimp. Furthermore, a number of genes encoding apoptotic-related proteins and antioxidant enzymes were expressed at a higher level in the virus-resistant shrimp. The high expression of the immune-relevant genes in response to the virus infection provides a new insight for further study in the shrimp innate immunity..