地高辛标记的金黄地鼠RPL30cDNA探针的制备和应用

目的制备地高辛标记的核糖体蛋白RPL30cDNA探针并用其研究RPL30在神经管发育过程中的表达状况。方法利用分部分排除技术（SSS法）从金黄地鼠神经管cDNA文库中逐级筛选得到RPL30cDNA的全长序列；随机引物法制备地高辛标记RPL30cDNA探针，点杂交法检测探针的灵敏度；Northern杂交检测神经管发育过程中RPL30mRNA的表达状况。结果本研究从构建的神经管cDNA文库中得到RPL30cDNA片段，全长535bp；制备探针的浓度达到50mg／L；Northern杂交检测发现E8d～E12d神经管中RPL30mRNA的表达均处于较高水平。结论该研究结果为进一步探讨RPL30与神经管发育和神经管畸形发生之间的关系奠定基础。

Preparation and Application of Dig-Labeled Rpl30cdna Probe

Objective To preparation a dig-labeled RPL30cDNA probe and use it to detect the level of RP- L30mRNA in the neural tube of golden hamster. Methods The RPL30eDNA segment was obtained from the neural tube cDNA library using subsection sereening method step by step;dig-labelled probe was prepared by random priming; and the level of RPL30mRNA in the neural tube was deteeted by Northern blot during the development of the neural tube. Results The RPL30eDNA was obtained from the neural tube eDNA library after screenings, and the length of the eDNA insert was 535bp, the concentration of the probe was 50mg/L. Northern blot showed that the level of RPL 30mRNA was high in the neural tube at each time fiom E8d - E12d. Conclusion Our resuhs may lay solid foundation for further researehing the relationship between RPL30 gene and the development of the neural tube and neural tube defects.