论文部分内容阅读
细胞因子在T 淋巴细胞胸腺内发育中起着重要的调节作用,然而生理条件下体内胸腺细胞发育过程中细胞因子基因表达的状态所知甚少。现首次报导应用简易敏感的PCR方法,扩增细胞因子的cDNA,结合半定量分析研究胸腺细胞发育中,诸细胞因子基因的表达水平。首先在6～12周龄定时配对后分笼饲养的基础上,分别获取第14～20天BALB/c 和C57BR10只以上胎鼠胸腺,抽提总RNA 进而转录成cDNA 作为模板,分别以人工合成的IL-1～IL-7,IFNγ,LT,GM-CSF,IL-2Rβ,β-肌球蛋白的特定序列为引物,PCR扩增15个循环后,每5个循环取样,共35个循环后行电泳分析或经杂交证明,同时以同
Cytokines play an important regulatory role in the thymus development of T lymphocytes, however, little is known about the state of cytokine gene expression during thymocyte development in vivo under physiological conditions. For the first time reported simple and sensitive PCR method, amplification of cytokine cDNA, combined with semi-quantitative analysis of thymocyte development, the expression of cytokine genes. First of all, on the basis of 6-12 week-old time-matched paired cage feeding, the thymus of BALB / c and C57BR10 fetuses of above fetus on day 14 to 20 were respectively obtained, total RNA was extracted and transcribed into cDNA as template, The specific sequences of IL-1 to IL-7, IFNγ, LT, GM-CSF, IL-2Rβ and β-myosin were used as primers. After 15 cycles of PCR amplification, samples were taken every 5 cycles for 35 cycles After the electrophoresis analysis or proven by hybrid at the same time with the same