斑节对虾GRP94基因的克隆及其在不同应激条件下的表达与分析

Characterization and expression analysis of GPP94 in black tiger shrimp(Penaeus monodon) under different acute stresses

以斑节对虾(Penaeus monodon)为研究对象,利用RACE技术克隆得到了斑节对虾Pm GRP94基因c DNA全长,并对其结构进行生物信息学分析。Pm GRP94全长2 990 bp,包括75 bp的5'UTR、527 bp的3'UTR和2 388bp的ORF。利用实时定量PCR技术,对Pm GRP94组织特异性及其在不同p H、盐度和3种重金属应激下转录水平变化情况及特点进行了研究。结果显示,Pm GRP94基因存在组织特异性,并在肝胰腺中的表达量最高;不同的应激条件下其表达具有明显差异,其中在p H、铜(Cu)和锌(Zn)的应激下Pm GRP94的表达显著升高。因此预测该基因可以作为斑节对虾受p H、Cu和Zn胁迫的指示基因。

In the present study,the full-length of Pm GRP94 was cloned from Penaeus monodon using RACE-PCR. The full-length of Pm GRP94 c DNA was 2 990 bp containing a 5＇ untranslated region（ UTR） of 75 bp,a 3＇ UTR of 527 bp and an open reading frame（ ORF） of 2 388 bp. We analyzed the characterization and expression,of tissue specificity of Pm GRP94 when the shrimps were exposed with different p H values,salinities and three heavy metals using real time PCR（ RT-PCR）. The highest expression level was found in hepatopancreas. The transcription levels of Pm GRP94 were significantly up-regulated under p H,copper and znic challenges.The results suggest that Pm GRP94 might be a promising candidate marker for p H stress, cadmium and manganese exposure in shrimps.