Panning for immune-related genes from T7 phage display library constructed using Chinese shrimp’s hemocyte RNA

LIU Ning; DU Xin-jun; ZHAO Xiao-fan; WANG Jin-xing

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LIU Ning, DU Xin-jun, ZHAO Xiao-fan, WANG Jin-xing. Panning for immune-related genes from T7 phage display library constructed using Chinese shrimp’s hemocyte RNA[J]. Haiyang Xuebao, 2007, 29(5): 102-108.

Citation:

LIU Ning, DU Xin-jun, ZHAO Xiao-fan, WANG Jin-xing. Panning for immune-related genes from T7 phage display library constructed using Chinese shrimp’s hemocyte RNA[J]. Haiyang Xuebao, 2007, 29(5): 102-108.

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Panning for immune-related genes from T7 phage display library constructed using Chinese shrimp’s hemocyte RNA

1.
School of Life Sciences Shandong University, Jinan 250100, China;Shandong Quangang Pharmaceutical Con Led Jinan 250014, China
2.
School of Life Sciences Shandong University, Jinan 250100, China

Received Date: 2006-03-08
Rev Recd Date: 2007-01-29

Abstract

Abstract

The objective was to find and identify immune-related genes of Chinese shrimp (Fenneropenaeus chinensis) by panning phage display library T7 phage display libraries were constructed with the shrimp hemocyte mRNA following the manufactures instruction.The titer of the Library was 1.1×10¹¹pfu/cm³.The library was panning by various ligands, including lipopoly saccharide, chitin and different kinds of bacteria, such as gram positive bacteria, Staphylococcusaureus, Bifidobacteriumlactis and Micrococcusluteus, and gramnegative bacteria, Escherichia coli, Vibrio anguillarum.After three rounds of phage panning, the inserts in the phage was amplified by PCR with T7 select 10-3b vector primers, and subcloned into pMD 18T vecter for sequencing.The sequences were analyzed by BLAST Sixteen fragments were obtained.Among them, peritrophin, Proteinarg inine N-methyltransferase, thr mbospondin, RNA binding protein, and nuclear autoantigenic sperm protein which were firstly reported in Chinese shrimp.The expression profile of peritrophin was analyzed by Reverse Transcription-PCR (RT-PCR), the results showed that it was reducing expression in hemocytes, heart, stomach, gut, gill and sperm in infected shrimp.This surgested that the peritriphin may be involved in innate immunity.Other genes panned from the library need further study.
- T7 phage display,
- innate immunity,
- pathogen associated molecular patterns,
- Chinese shrimp

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References(15)

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