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目的:研究nfic基因在牙齿发育中的作用。方法:提取出生后SD大鼠磨牙牙胚组织总mRNA, 反转录合成cDNA。根据NCBIGeneBank中nficcDNA序列,分别设计2对PCR引物,进行nfic基因部分序列的扩增,扩增产物与T载体进行T/A连接,然后转化高效感受态大肠杆菌DH5α,挑取阳性克隆进行酶切鉴定,并对扩增产物进行序列测定。结果:P1、P2为引物进行PCR扩增,可见大小约350bp的特异扩增产物;以P3、P4为引物进行PCR扩增,电泳可见大小不同的3条特异扩增产物,经克隆后的序列测定,为大鼠nfic基因的3种可变剪接体。结论:nfic基因在大鼠牙齿发育后期、牙根的发育前期有表达,并且存在至少3种大鼠nfic基因选择性剪接体。
Objective: To study the role of nfic gene in tooth development. Methods: Total mRNA of tooth germ tissue of SD rats was extracted and cDNA was reverse transcribed. According to the sequence of nfic cDNA in NCBIGeneBank, two pairs of PCR primers were designed respectively to amplify the nfic gene partial sequence. The amplified product was T / A linked to T vector and then transformed into competent E. coli DH5α. The positive clones were selected for digestion Identification, and amplification products were sequenced. Results: P1 and P2 were used as primers for PCR amplification, and the specific amplification products were about 350bp in size. Three specific amplification products with different sizes were obtained by PCR using P3 and P4 as primers. The cloned sequences Determination of the rat nfic gene 3 alternative splicing. CONCLUSION: The nfic gene is expressed in the early period of tooth development and root development in rats, and there are at least 3 alternative splicing of nfic gene in rats.