Construction Of T₇ Phage Display Library From Chinese Shrimp's Hemocytes And Panning Of Functional Genes From The Library

With the development of shrimp aquaculture, shrimp diseases are becoming more and more frequently and seriously. Nowadays the immunology mechanism of shrimp is not well understood, so there is little valid measures to prevent and treat certain diseases which leading to great economic losses. To investigate immune-related genes and immunology mechanism of shrimp, such as pattern recognition receptors, we constructed T₇ Phage display library from Chinese shrimp's hemocytes. Panning the library with different ligands and methods , we gained some valuable genes and then taken a further research. The data and results of my paper can provide us with basic information of shrimp's immunological mechanism. Hereunder are results of my research:1. T₇ Phage Display Library was constructed from challenged shrimp's hemocyte mRNA, and then the library was amplified to reach the titer of 10¹¹pfu/ml.2. The Library was panned by various ligands, including lipopolysaccharide, chitin and different kinds of bacteria. After phage panning, fragments subcloning and sequencing, sixteen gene fragments were obtained, Among them peritrophin, arginine N-methyltransfer, calreticulin, RNA binding protein and Nuclear autoantigenic sperm protein which were firstly reported in shrimp.3. Further reseach was done to analysis RNA binding protein which was panned with Micrococcus luteus. The expression and distribution of this gene in different tissues were analyzed by RT-PCR, Northern Blot and in situ hybridization. The results revealed that RNA binding protein gene highly expressed in ovary and the expression of this gene was up-regulated in hemocytes, heart, hepatopancreas and stomach after challenged with bacteria.4. Elongation factor 1-α was panned with LPS and the full length of the gene was...