The Construction of Intercellular Adhesion Molecule-1 cDNA Expression Vector / 中国医师杂志

ABSTRACT

Objective To construct ICAM-1 recombinant eukaryotic expression vector. Methods Human intercellular adhesion molecule-1 (ICAM-1) cDNA was obtained by RT-PCR of totol RNA extracted from human hepatocellular carcinoma tissue. Amplified ICAM-1 cDNA fragment was cloned into pGEM-T easy vector to construct pGEM-ICAM-1 vector. Then ICAM-1 cDNA from pGEM-ICAM-1 vector was cloned into eukaryotic expression pcDNA3.1hisB to construct recombinant pcDNA3.1hisB-ICAM-1 vector. Restriction endonuclease digestion and DNA sequencing were used to confirm the recombinant vector. Results 1622bp ICAM-1 cDNA was obtained by RT-PCR. The PCR product was successfully ligated with pGEM-I easy vector. Restriction endonuclease digestion analysis and DNA sequencing showed that recombinant pcDNA3.1HisB-ICAM-1 was successfully constructed. Conclusion Eukaryotic expression recombinant vector pCDNA3.1hisB-ICAM-1 was contructed.