小鼠PeriostinC端编码区cDNA克隆及其序列分析

目的：从小鼠牙周膜组织中克隆PerostinC末端编码区基因。方法：用异硫氰酸胍一步法从昆明成年小鼠牙周膜组织中抽提总RNA，用Oligo(dt)作引物逆转录合成cDNA，然后利用PCR技术，从cDNA中扩增出小鼠PeriostinC末端编码区的基因自然（约940bp），将所得基因片段插入p RSET-B载体，转化大肠杆菌DH5α后随机挑选数个克隆，提取质粒DNA，通过限制性酶切和核苷酸序列分析鉴定阳性克隆。结果：重组质粒pRSET-B-Periostin的酶切图谱和序列分析结果与国外文献报道一致。结论：克隆到小鼠该蛋白C端编码区基因。

Cloning and sequencing of the terminal C encoding gene of mouse Periostin

AIM: To get the terminal C encoding gene of Periostin from mouse periodontium. METHODS: Total RNA was extracted from the periodontium of adult mouse by acid guanidinium thiocyanata-phenol-chloroform method. The desired DNA product was obtained from the total RNA by RT-PCR with the primers including Oligo(dt) and two gene specific primers. The segment (about 940bp) was inserted into pRSET-B vector and the insered plasmid was transformed into E.coli DH5α.The positive clone was analyzed by restriction endonuclease mapping and DNA sequencing. RESULTS: The restriction endonuclease map and sequence of mouse Periostin functional fragment were consistent with those of the published. CONCLUSION: The terminal C encoding gene of mouse Periostin has been cloned.