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目的:克隆小鼠IL-33基因全长编码区cDNA,并对其进行序列分析。方法:从BALB/c小鼠的脊髓组织中提取总RNA,逆转录为cDNA,用热启动PCR技术,扩增小鼠IL-33基因全长编码区cDNA,经双酶切后,克隆入pcDNA3.1(+)载体中,构建真核表达载体pcDNA3.1-mIL-33,然后进行酶切鉴定与序列分析。结果:小鼠IL-33基因的PCR产物和重组载体经凝胶电泳和酶切鉴定、测序分析证实,其序列与GenBank中数据一致。小鼠IL-33基因的全长编码序列为801 bp,编码266个氨基酸。结论:小鼠IL-33基因成功的克隆并构建了其真核表达载体,为进一步进行IL-33的表达与功能研究奠定了基础。
OBJECTIVE: To clone the full length coding region of mouse IL-33 gene and analyze its sequence. METHODS: Total RNA was extracted from the spinal cord of BALB / c mice and reverse transcribed into cDNA. The full-length coding region of mouse IL-33 gene was amplified by hot-start PCR and cloned into pcDNA3 .1 (+) vector, construct eukaryotic expression vector pcDNA3.1-mIL-33, and then carry on restriction enzyme digestion and sequence analysis. Results: The PCR product and recombinant vector of mouse IL-33 gene were identified by gel electrophoresis and restriction enzyme digestion. Sequencing analysis confirmed that the sequence of the IL-33 gene was consistent with that of GenBank. The full-length coding sequence of mouse IL-33 gene is 801 bp, encoding 266 amino acids. Conclusion: The mouse IL-33 gene was successfully cloned and its eukaryotic expression vector was constructed, which laid the foundation for the further study on the expression and function of IL-33.