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Updated: Aug 7, 2026
The Multifaceted Benefits of Protein Co-expression in Escherichia coli
Published on: February 5, 2015
[Construction of mammalian co-expression plasmid pIRES -LMP1-HSP90 beta and its expression in vitro]
Ju-mei Zhao1, Tao Liu, Ling Tian
1State Key Laboratory of Biotherapy Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
Objective:
This study sought to clone Epstein-Barr virus latent membrane protein 1 (LMP1) and heat shock protein 90 beta (HSP90 beta) of nasopharyngeal carcinoma (NPC), to construct the mammalian co-expression plasmid pIRES-LMP1-HSP90 beta, and to detect the expression of the plasmid in vitro.
Methods:
Total RNA was isolated from human NPC by cloning technique, their cDNA fragments of LMP1 gene and HSP90 beta gene were gained by RT-PCR, and their cDNA fragments were constructed into the mammalian co-expression plasmid vector pIRES. The inserted target genes in the mammalian co-expression plasmid were verified by nucleotide sequencing. COS cell line was transfected with this mammalian co-expression plasmid using lipofectin reagent. The expression of LMP1 and HSP90 beta molecules were detected by RT-PCR and Western blot technique.
Results:
The mammalian expression plasmid pIRES-LMP1-HSP90 beta was obtained by cloning technique. The nucleotide sequences of LMP1 gene and HSP90 beta gene in this mammalian co-expression plasmid had high homology with EBV-LMP1 (100%) and human HSP90 beta (100%) respectively. After transfection with this mammalian co-expression plasmid, the LMP1 and HSP90 beta molecules were expressed in COS cells.
Conclusion:
The constructed mammalian co-expression plasmid pIRES-LMP1-HSP90 beta can express LMP1 and HSP90 beta molecules in vitro at the same time.
