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中文摘要：
目的：制备NIDD基因TaqMan探针建立检测NIDD基因荧光定量PCR（FQ-PCR），以进一步研究NIDD的表达情况。方法：采用RT-PCR法，从出生后一天的SD大鼠脑组织mRNA中扩增NIDD基因的部分编码序列区（CDS）片段，克隆入pGEM-T载体，筛选阳性克隆、酶切鉴定及序列测定。采用TaqMan探针，以重组质粒为模板绘制标准曲线。结果：RT—PCR法扩增出一特异产物与预期长度112bp相符，DNA序列测序的结果与C,eneBank提供的已知序列（AB098078）比较，所克隆的NIDD基因片段与其中的112bp完全相同，与NIDD基因100％同源。以重组质粒为模板绘制标准曲线，相关系数r大于0．99，反应效率显示为1，各点变异系数小于10％。结论：采用RT．PCR和T载体技术成功制备出可应用于FQ-PCR的NIDD基因TaqMan探针。
英文摘要：
Objective： To obtain the probe of a novel nNOS-interacting DHHC domain-containing protein with dendritic mRNA （NIDD） for real-time fluorescent quantitative PCR （FQ-PCR）. Methods： The partial CDS of NIDD was amplified by RT-PCR from ld postnatal SD rat brain. RT-PCR product was ligated into pGEM-T vector, and the DNA sequence was detected. The standard curve was drawn with the recombinant plasmids and the NIDD TaqMan probe. Results： The product of RT-PCR was 112 bp which matched the size of the purpose. This DNA sequence was 100% homogeneous with the NIDD cDNA reported. The correlation coefficient of the standard curve was above 0.99, the reaction efficiency was 1 and all the variation coefficients were below 10%. Conclusion： The NIDD TaqMan probe for FQ-PCR has been successfully obtained with RT-PCR and T vector techniques.
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