大黄鱼Hepcidin基因转录物的相对定量分析

以大黄鱼(Pseudosciaena crocea)管家基因18S rRNA和β-actin作为内参基因,分别比较2个内参基因建立的相对定量曲线,最终确立以18S rRNA为参比基因,定量分析大黄鱼的Hepcidin抗菌肽基因.该相对定量分析方法所得结果与Northern-blot方法一致.应用建立的Hepcidin基因的实时荧光相对定量研究方法,对大黄鱼头肾中的Hepcidin基因转录物进行相对定量,为今后开展鱼体内免疫相关基因的表达特性、诱导机制等工作奠定研究基础.同时,克隆得到的大黄鱼β-actin基因和18S rRNA基因片段已提交基因库,并获得登录号.

Relative Quantification of Hepcidin Gene Transcripts in Pseudosciaena crocea

Two house-keeping genes,18S rRNA and β-actin,were cloned as endogenous genes for relative quantification of an antimicrobial peptide hepcidin in large yellow croakers(Pseudosciaena crocea) using the real-time RT-PCR.In comparision of the slopes of the relative standard curves between the two endogenous genes and the target gene,a relative quantification method for hepcidin gene using 18S rRNA as the calibrator gene was established.Results obtained with this method were consistent with the rusults from Northern-blot,suggesting feasibility and effectiveness of the method.This paper provides an effective method to investigate expression patterns and induction profiles of the hepcidin gene in large yellow croakers,and other immunity-related genes in fish bodies.The cloned fragments of 18S rRNA and β-actin genes from the large yellow croaker have been submitted to the GenBank,which have provided accession numbers.