家蚕Gar1p基因的克隆表达及生物信息学分析

Cloning and Bioinformatic Analyses of Gar1p Gene From Bombyx mori

在对家蚕功能基因筛选中获得一条长844 bp的基因序列,其编码蛋白与一种富含甘氨酸和精氨酸的核仁小分子RNA结合蛋白(glycine and arginine-rich nucleolar protein,Gar1p)的同源性达63%。利用生物信息学方法分析了该基因上游的启动子序列,发现该基因不含内含子,由1个699 bp的单一外显子编码232个残基的蛋白,预测分子量为22.4 kD,等电点11.43。该Gar1p蛋白的疏水性最大值为1.167,最小值为-2.011,其序列的两端各有1个强亲水性的GAR区域,而中间含有5个潜在的磷酸化位点。Gar1p基因已从家蚕基因组中扩增得到,并克隆进表达载体pET-28a中,在大肠杆菌中得以表达,为研究其功能提供了条件。

In scanning for functional genes from gene library of Bombyx mori, an 844 bp gene was identified, which encodes a protein of homogenous with a glycine and arginine-rich nucleolar protein Garl p. Ther homogeneity reaches 63%. By means of bioinformatic methods, the promoter in upstream of this Gar lp gene was detected. This gene contains no intron but an exon of 699 bp encoding 232 amino acids, with a predicted molecular weight of 22.4 kD and pl of 11.43. The hydrophobicity of this Garl p protein was predicted to be 1. 167 in maximum and -2. 011 in minimum. Each terminal of the protein contains a hydrophilic GAR domain, while the middle of the protein has 5 potential phosphorylation sites. This Gar lp gene was amplified from genomic DNA of Bombyx mori, cloned into pET-28a vector and successfully expressed in E. coil, supporting for functional studies of this gene.