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目的建立SMP1基因外显子检测方法以探讨SMP1基因多态性。方法根据文献报道并参考Gene-Bank中的基因序列,设计聚合酶链式反应(PCR)扩增SMP1基因7个外显子的8对特异性引物,应用PCR分段扩增SMP1基因7个外显子,PCR产物纯化后直接测序,应用DNAMAN5.2.9序列分析软件分析序列。结果 SMP1基因第1~7外显子长度分别为137、106、113、68、93、60和1 703 bp。SMP1基因第1~6外显子未发现多态性位点,但该基因第7外显子存在1329T>C和1704G>A多态性。结论本研究成功建立SMP1基因外显子检测方法并检测到了SMP1基因第7外显子存在多态性位点。
Objective To establish a method for exon detection of SMP1 gene to investigate the polymorphism of SMP1 gene. Methods Eight pairs of primers specific to the seven exons of SMP1 gene were designed by polymerase chain reaction (PCR) based on the reported gene sequences in Gene-Bank. Seven fragments of SMP1 gene were amplified by PCR Exons, PCR products were purified directly sequenced DNAMAN5.2.9 sequence analysis software analysis of the sequence. Results The lengths of exon 1 to exon 7 of SMP1 gene were 137, 106, 113, 68, 93, 60 and 1 703 bp, respectively. Polymorphic sites were not found in exon 1 to exon 6 of SMP1 gene, but there were 1329T> C and 1704G> A polymorphisms in exon 7 of this gene. Conclusion This study successfully established the exon detection method of SMP1 gene and detected the polymorphism of exon 7 of SMP1 gene.