鹅CD4与CD8α实时荧光定量PCR检测方法的建立
Quantification of Chinese goose CD4 and CD8α mRNA using real-time reverse transcription polymerase chain reaction assay
4+ T又被称作辅助性T淋巴细胞，CD8+ T又称细胞毒性T淋巴细胞。在细胞免疫应答中都发挥很重要的作用。在本文中，根据我们前期获得的四川白鹅CD4、CD8α的cDNA序列，设计并合成特异性引物，建立了一种能够实时检测鹅CD4、CD8α分子转录水平的荧光定量PCR检测方法。两个基因的标准曲线R2大于0.99，表明线性度良好，扩增效率也都高于90%，证明建立的检测方法是可行的。在此基础上，利用分离的鹅脾脏淋巴细胞作为体外试验的细胞模型，用我们建立方法来检测鹅CD4、CD8α分子在体外实验模型中的生物学活性，研究发现PHA可以诱导CD4+、CD8+T细胞的活化。本检测方法的建立为进一步研究鹅CD4与 CD8α分子的生物学活性以及免疫学活性的奠定了基础。
4+ T cells, or helper T cells, and CD8+ T cells, or killer T-cells, execute different aspects of T cell mediated adaptive immunity. Several specific primers were designed according to the cDNA sequences of goose CD4 and CD8α, and the goose β-actin was used as internal control. The correlation coefficient (R2) values of the standard curves for all target genes were above 0.99, and qPCR efficiency for all target genes were all above 90% .In this study, the rapid and specific real-time qPCR methods to analyze the transcription profiling of goose CD4 and CD8α gene were established, and the specific real-time qPCR methods were performed for the detection of the biological activity of goose CD4 and CD8α in the spleen mononuclear cells (MNCs). The methods we established here make it available to further research on immunological activity of goose CD4 and CD8α.
赵秋荣、陈舜
分子生物学细胞生物学
四川白鹅48α荧光定量PCR
Sichuan white gooseCD4CD8αreal-time qPCR
赵秋荣,陈舜.鹅CD4与CD8α实时荧光定量PCR检测方法的建立[EB/OL].(2013-11-15)[2025-07-16].http://www.paper.edu.cn/releasepaper/content/201311-295.点此复制
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