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Updated: Jul 13, 2026
Optimization of a Multiplex RNA-based Expression Assay Using Breast Cancer Archival Material
Published on: August 1, 2018
[Expression of (His)(6)-eIF3s4 fusion protein in human breast cancer cell Bcap37]
Qun Wei1, Feng-jia Zhu, Yi Wang
1Sir Run Run Shaw Hospital Clicinal Research Institute, College of Medicine, Zhejiang University, Hangzhou 310016, China.
Aim:
To construct eukaryotic expression vector of eukaryotic translation initiation factor 3 subunit 4(eIF3s4) for further functional study.
Methods:
The cDNA of eIF3s4 containing full length coding region was amplified by RT-PCR and cloned into pGEM-T Easy. The sequence of the cDNA was verified by DNA sequencing and blast against sequence data in GenBank database. The cDNA was then subcloned into the pcDNA4/HisMaxB to make a (His)(6)-eIF3s4 fusion protein expression vector. The vector was transfected into human breast cancer cell Bcap37 by Lipofectamine 2000, and the expression of (His)(6)-eIF3s4 fusion protein was detected by Western blot.
Results:
DNA sequencing and sequence blast showed that the cDNA amplified by RT-PCR was consistent with the eIF3s4 sequence in GenBank database, and Western blot results showed the expression of the (His)(6)-eIF3s4 fusion protein in human breast cancer cell Bcap37 as expected.
Conclusion:
The (His)(6)-eIF3s4 fusion protein expression vector is constructed successfully with expression of the fusion protein in Bcap37 breast cancer cells. This work provides the basis for establishing a stable eIF3s4 expressing cell line for further study on the role of eIF3s4 in cancer multidrug resistance.
