Feasibility study of bacterial lipopolysaccharides to increase black tiger shrimp survival by using an array-based method to monitor shrimp immune system
Aim 1: To develop multiplex immune gene expression analysis using bead array platform. The oligonucleotide probes will be designed to target immune-related genes of P. monodon. The gene specific probe will be conjugated to a carboxylated magnetic bead, and fluorescent signal will be detected in Bio-plex bead array system. The hybridization conditions will be optimized to obtain high fluorescent signal intensity. The specificity and sensitivity of each probe will be determined using complementary oligonucleotide and plasmid DNA templates.
Aim 2: To validate multiplex immune gene expression array. The multiplex array will be applied to compare expression of target genes using RNA extracted from P. monodon tissues under pathogen exposure. The gene expression profiles will be compared to results obtained from realtime PCR assay.
Aim 3: To assess efficacy of immune stimulation effects in the black tiger shrimp by lipopolysaccharides obtained from different bacterial sources. Survival rates of P. monodon will be compared under pathogen exposure when given LPS from different bacterial sources (native and non-native to shrimp intestine). Shrimp immune system will be determined at a molecular level.
