梅花鹿成熟型TGF-β1基因的克隆与结构分析及其在鹿茸顶端组织中的表达

Cloning and Structural Analysis of Mature Type TGF-β1 Gene in Sika Deer and Its Expression in Antler Tissue

为探究细胞因子对鹿茸再生的影响,提取鹿茸顶端组织总RNA(Trizol法),反转录获得cDNA模板,克隆TGF-β1成熟肽基因。利用生物信息学软件,对其基因序列及蛋白质结构进行分析。结果表明:成功获得梅花鹿TGF-β1成熟肽基因,序列长339 bp,共编码112个氨基酸,蛋白性质稳定,理论分子质量约为12.8 kD。同源性分析结果显示,TGF-β1在进化过程中具有高度保守性,梅花鹿与牛、羊、猪、人、鼠的核苷酸序列同源性均>90%;除鼠外,梅花鹿与其他物种氨基酸序列并未发生变化。免疫组化结果显示,TGF-β1在鹿茸顶端不同组织中差异表达,软骨层中表达水平最高。由此推测,TGF-β1在鹿茸再生过程中可能发挥重要作用。

To explore the effects of cytokines on antler regeneration, the total RNA from the tip tissue of antler was extracted and cDNA template was obtained by reverse transcription. The TGF-β1 mature peptide gene was cloned by RT-PCR technology. Then, bioinformatics software was used to analyze the sequence and structure of the TGF-β1 gene. The results showed that the mature peptide gene of TGF-β1 has been successfully obtained. It has 339 base pairs in length, encoding a protein of 112 amino acids. The encoded protein has stable physical and chemical properties, and the molecular weight is about 12.8 kD. The results of homology analysis showed that TGF-β1 is highly conserved in the evolution process, and the nucleotide sequence homology of sika deer with cattle, sheep, pig, human and mouse is more than 90%. The amino acid sequence of sika deer and other species does not change except in rat. The results of immunohistochemistry showed that TGF-β1 is differentially expressed in different tissues on the top of antler, and the expression level of TGF-β1 is the highest in the cartilage layer. It is speculated that TGF-β1 may play an important role in the process of antler regeneration.