藏山羊GEM基因克隆及组织表达分析

Cloning and Tissue Expression Analysis of GEM Gene in Tibetan Goat

本研究旨在克隆获得藏山羊GEM基因序列,预测其生物学功能,并阐明其组织表达特征。利用TA法克隆获得藏山羊GEM基因序列,并利用qPCR技术检测该基因在不同组织中的表达丰度。结果显示,藏山羊GEM基因序列1 096 bp,CDS区为936 bp,共编码311个氨基酸;GEM蛋白具有磷酸化位点和糖基化位点共33个,为不稳定碱性疏水蛋白质,主要在细胞核发挥生物学作用;藏山羊GEM基因的核苷酸序列和氨基酸序列均与反刍动物的序列具有很高的同源性;GEM基因在藏山羊肺组织中高表达(p<0.01),其次高表达于脾组织(p<0.01),均极显著高于其他组织。本研究为进一步了解藏山羊GEM基因功能提供基础数据。

The aim of this study was to obtain the GEM gene sequence of Tibetan goat by cloning,forecast its biological functions and illuminate its expression characteristics in different tissues.The GEM gene sequence of Tibetan goat was cloned and obtained by TA method,the expression levels of GEM gene in different tissues was detected by quantitative real-time PCR(qPCR).The results showed that the length of GEM gene sequence of Tibetan goat was 1 096 bp and the CDS region was 936 bp,encoding 311 amino acids.The GEM protein had 33 phosphorylation and glycosylation sites,which belonged to unstable alkaline hydrophobic protein,playing a biological role primarily in the nucleus.The nucleotide and amino acid sequences of Tibetan goat GEM gene were highly homologous to those of ruminants.The expression level of GEM gene in lung of Tibetan goat was the highest(p<0.01),and the second highest expression was in the spleen(p<0.01),which were significantly higher than that of other tissues.This study could provide basic data for the future study of GEM gene functions in Tibetan goat.