柞蚕核糖体蛋白基因S3a的克隆及表达分析

Cloning and Expression Analysis of Ribosomal Protein S3a Gene from Antheraea pernyi

核糖体蛋白在蛋白质的生物合成、细胞的代谢与凋亡、机体免疫、信号转导等方面有重要作用。采用RT-PCR方法克隆了柞蚕(Antheraea pernyi)核糖体蛋白基因S3a的开放阅读框(ORF),ORF序列长795bp,编码264个氨基酸。序列比对表明,柞蚕S3a蛋白与其它10个物种S3a蛋白的相似性介于72%～99%之间,其中与柳蚕(Actias selene)S3a蛋白的相似性最高。用RT-PCR方法分析该基因在柞蚕幼虫组织中的表达情况,结果显示柞蚕S3a基因在柞蚕幼虫的血液、中肠、丝腺和脂肪体组织中均有表达,且转录水平无显著差异。SDS-PAGE和Westernblotting检测显示柞蚕S3a基因在大肠杆菌中获得了正确表达。

Ribosomal proteins play important roles in protein biosynthesis, cellular metabolism, cell apoptosis, organismal immunity and signal transduction etc. In present study, the ribosomal protein S3a gene was cloned by RT-PCR from Chinese oak silkworm, Antheraea pernyi. Its ORF consists of 795 bp, encoding a protein with 264 amino acids. Sequence alignment revealed that A. pernyi S3a protein is 72% to 99% identical with those from other 10 species, among which Actias selene has the highest sequence identity with A. pernyi. RT-PCR analysis showed that the A. pernyi S3a gene was extensively expressed in hemocyte, midgut, silk gland and fat body, and the transcriptional level was not significantly different. The results of SDS-PAGE and Western blotting showed that the recombinant A. pernyi S3a protein was correctly expressed in Escherichia coll.