构建人ICAM-1真核表达载体的实验研究

CONSTRUCTION AND EXPRESSION OF HUMAN INTERCELLULAR ADHESION MOLECULE-1 EXPRESSION VECTOR

目的:构建人ICAM-1-1真核表达载体pcDNA3.1(-)-ICAM-1。方法:设计特异性引物,利用PCR技术扩增出ICAM-1全编码区基因片段,插入到真核表达载体pcDNA3.1(-)中。结果:PCR扩增得到人成熟ICAM-1的cDNA片段大小为1800bp,重组子利用限制性内切酶进行酶切鉴定和DNA序列分析得到真核表达载体pCDA3.1(-)-ICAM-1。结论:成功构建了人ICAM-1真核表达载体,为进一步建立稳定转染ICAM-1的细胞株以及研究ICAM-1及其受体的生物学功能提供了条件。

To construct ICAM - 1 eukaryotic expression vectors. Methods：ICAM - 1 cDNA was amplified by PCR and then inserted into the pcDNA 3.1 vector. Results ：1800 bp ICAM -1 cDNA was obtained by PCR. The PCR product was successfully ligated with pcDNA 3.1 vector. Restriction endonuclease digestion analysis and DNA sequencing showed that recombinant pcDNA 3.1 - ICAM - 1 was successfully constructed. Conclusion： Eukaryotic expression recombinant vector pcDNA 3.1 - ICAM - 1 was contructed, which lays the foundation for functional research of ICAM - 1 and for preparation of mAb to ICAM - 1.