Keywords

Interferon, Transmembrane Protein-1 Gene, Colorectal Tumors, Pcr, Cloning, Protein Expression

Abstract

Objective To study the amplification, cloning and protein expression of interferon-inducible transmembraneprotein-1 (ifitmp-1) gene. Methods The cdna containing ifitmp-1 gene was used as a template, PCR amplification was performed with pfu enzyme, and the target gene was cloned into pucm-t plasmid for sequencing using ecor¢¡ and hind¢£ double enzyme digestion. Further clone into the pet-trx protein expression vector plasmid and optimize the protein expression conditions. The result was that the PCR product containing ifitmp-1 gene was about 1000bp. The recombinant pucm-t plasmid was digested by ecor¢¡ and hind¢£, and a cdna fragment of corresponding size was inserted. The sequencing results showed that the sequence was correct, confirming that the target gene was successfully amplified and cloned, and was successfully cloned into the pet-trx protein expression vector. Induced by iptg, the fusion protein is expressed in bl21(de3)plyss. Conclusion The ifitmp-1 gene was successfully amplified and cloned, and the protein encoded by the gene was successfully expressed, which laid the foundation for further research on the role of ifitmp-1 gene in colorectal cancer.

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Original research done by Liu Yuhu, Zhong Dong, Liu Juan, Zhang Zhenshu, Chen Cunlong, Wu Jinbao, Xiao Bing, Guo Wenying

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