鸭Rab6基因cDNA的克隆及组织特异性表达

cDNA Cloning and Tissue Specific Expression of Rab6 Gene in Duck

【目的】研究鸭Rab6基因序列并进行序列分析,揭示该基因的组织特异性表达规律。【方法】运用RT-PCR技术获得鸭Rab6基因序列,并用半定量RT-PCR方法研究该基因的组织特异性表达规律。【结果】从35周龄母鸭脂肪组织中克隆了Rab6基因的cDNA序列,大小为761bp,包含1个627bp完整开放阅读框,编码208个氨基酸。该序列与小鼠、狗、人等物种的Rab6有86.1%～93.1%的同源性,而相应蛋白的同源性达97.6%以上。蛋白预测的分子量为23534D,等电点预测为5.42,该蛋白在78～102氨基酸处有1个跨膜结构,这些特征与人、小鼠、狗等物种高度相似。半定量RT-PCR研究结果显示,Rab6基因在所测组织中均表达,在小肠、肝、脾、间脑、肾、脂肪组织中高度表达,腺胃、骨骼肌、肌胃组织中中度表达,心、肺、卵巢中低度表达。【结论】Rab6基因在动物进化中具有高度保守性,具有相似的生物学功能,该基因的表达具有组织特异性。

[Objective] This study was carried out to obtain and analyze sequence of duck Rab6 gene, and to reveal the expression of Rab6 mRNA in duck different tissues. [Method] The sequence of duck Rab6 gene was obtained by reverse transcription PCR （RT-PCR） and its expression in different tissues was studied by semi-quantity RT-PCR. [ Result ] A 761 bp cDNA ofRab6 gene was cloned and sequenced from the adipose tissue of 35-week-old female duck. It contains a complete 627 nucleotides open reading-fragment （ORF）, encoding 208 amino acids. The duck Rab6 CDS shares 86.1%-93.1% homology with Rab6 CDS of house mouse, dog, human, and so on, while the corresponding protein shares more than 97.6% homology with the above animals＇ Rab6 protein. The predicted molecular weight （MW） and isoelectric point （pI） of the protein are 23 534 D and 5.42, respectively, and it has a transmembrane domains at 78-102 amino acid, which all are highly similar to the above animals＇. The semi-quantity RT-PCR revealed duck Rab6 mRNA displayed high expression levels in small intestine, liver, spleen, diencephalons, kidney, adipose, middle expression in glandular stomach, skeletal muscle, muscular stomach, and weak expression in lung, heart and ovary. [Conclusion] Rab6 gene is of high conservation in the course of animal evolution, and they have approximate biological function in different animals. The expression level ofRab6 mRNA is particular to tissues.