Genes Expression in Hemocytes of the Giant Freshwater Prawn (Macrobrachium rosenbergii) Elicited by CpG Oligodexoynucleotide by Suppression Subtractive Hybridization

利用抑制性扣減雜交作用測定CpG寡核苷酸誘發之淡水長腳大蝦(Macrobrachium rosenbergii)血球的基因表現

路光予(Kaun-Yu Lu) ； 宋昕如(Hsin-Ju Sung) ； 宋宏紅(Hung-Hung Sung)

《臺灣水產學會刊》 34卷3期 (2007/09) Pp. 275-290

https://doi.org/10.29822/JFST.200709.0004

摘要

為了瞭解蝦類的免疫反應，許多免疫相關的蛋白質和其基因曾被分離出來，並對它們的活性和免疫角色進行研究。本研究以CpG寡核苷酸(ODN)誘發淡水長腳大蝦後，針對誘發和未誘發兩組的蝦子血球之基因表現模式作系統性分析。每隻蝦子注射5μg的ODN2006後1、6、及12小時，採取蝦血球，製備RNA樣本；接著，利用抑制性扣減雜交(SSH)技術鑑定ODN注射組和未注射組蝦子血球之基因表現模式的差異。差異表現的基因主要可以分為兩群：受正調控的基因群（表現增加的基因）和受負調控的基因群（表現受到抑制的基因）。根據生理功能，受正調控的基因群可以進一步的分為不同的五類，包括防禦相關分子的基因、蛋白質合成及折疊相關的基因、產生能量系統的基因、不知功能的基因、以及其他。依據基因表現的時間先後，推測CpG ODN刺激的蝦子之免疫反應具有－系統性基因表現的模式。

關鍵字

系統性基因表現 ； CpG寡核苷酸ODN ； 蝦類 ； 抑制性扣減雜交

並列摘要

In order to understand the immune response of prawns, different kinds of immune-related proteins and their genes had been isolated, their activities and their possible immunological roles had been investigated. In this study, a systematic analysis of the gene expression pattern between CpG oligodeoxynucleotide (ODN)-elicited and -unelicited prawn Macrobranchium rosenbergii was undertaken. The RNA of hemocytes was extracted from the prawns at 1, 6 and 12 hours after prawn injected with 5μg/prawn of ODN2006. The difference of gene expression patterns between the hemocytes of ODN-treated and untreated prawns were identified by using the technique of suppression subtractive hybridization (SSH). Among those differentially expressed genes, two major categories of genes were identified: up-regulated genes (gene expression has been elevated) and down-regulated genes (gene expression has been suppressed). The up-regulated genes could be further classified into 5 different kinds according to their physiological functions, including: genes concerning the defense-related molecules, genes concerning the protein synthesis and folding, genes concerning the energy producing system, and those genes concerning till now unknown functions and others. According to the time course of the gene expression, it is suggested that a systematic gene expression pattern exists in the immune response of prawns stimulated by ODN2006.

並列關鍵字

Systematic gene expression ； CpG Oligodeoxynucleotide ODN ； Prawn ； Suppression subtractive hybridization SSH