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目的:克隆非人灵长类疾病动物模型-恒河猴的CD1d基因并检测其在不同组织中的表达差异情况。方法:提取恒河猴血液及各组织的RNA,以反转录得到的cDNA为模板,用特异性引物扩增CD1d;以管家基因GAPDH为内参,用半定量RT-PCR的方法对CD1d的组织表达差异性进行分析。结果:成功扩增了恒河猴CD1d基因的编码区序列;首次用半定量RT-PCR的方法检测了CD1d mRNA在恒河猴部分组织中的表达情况,发现其表达水平存在明显的组织差异性,其中,在肝脏、脾脏和心脏中的表达水平较高,在血液、小肠和肺中次之,在脑中表达最少。结论:研究结果为今后表达恒河猴CD1d蛋白并制备其四聚体进而研究恒河猴NKT细胞在众多疾病中的作用奠定了基础;组织表达差异的研究结果提示其在相关疾病的发生发展过程中可能扮演着重要的角色,具体作用还有待深入研究。
OBJECTIVE: To clone the CD1d gene of rhesus monkeys, an animal model of non-human primate diseases, and to detect the difference in expression among different tissues. Methods: RNA was extracted from the blood and tissues of rhesus monkeys. Reverse transcribed cDNA was used as a template to amplify CD1d with specific primers. The housekeeping gene GAPDH was used as an internal control. The expression of CD1d Expression differences were analyzed. Results: The coding region of CD1d gene of Rhesus monkey was successfully amplified. The expression of CD1d mRNA in some rhesus monkeys was detected by semi-quantitative RT-PCR for the first time and found that there was significant difference Among them, the expression level in the liver, spleen and heart is higher, secondarily in the blood, small intestine and lung, and the least in the brain. Conclusion: The results of this study lay the foundation for future expression of rhesus monkey CD1d protein and preparation of its tetramer and further study of the role of rhesus NKT cells in many diseases. The results of tissue expression suggest that it may play an important role in the development of related diseases May play an important role in the specific role remains to be further studied.