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Updated: Jul 18, 2026
Spectrophotometric Screening for Potential Inhibitors of Cytosolic Glutathione S-Transferases
Published on: October 10, 2020
[Expression of glutathione-S-transferase fusion protein and human CCL3L1 protein]
Bin Xu1, Ying Shi, Jun-Hong Li
1Department of Infectious Diseases, Beijing Youan Hospital, Capital University of Medical Sciences, Beijing 100069, China.
Objective:
To clone human CCL3L1 cDNA and to express and purify the glutathione-S-transferase (GST) fusion protein and human CCL3L1 protein.
Methods:
Total RNA was isolated from breast cancer cell line MCF7. CCL3L1 cDNA including open reading frame was obtained by RT-PCR. PCR product was digested with EcoR I and cloned into the pGEX-4T-1 vector. The plasmids from positive clone was prepared and sequenced to confirm the CCL3L1 in correct fusion form. pGEX-4T-CCL3L1 was transfected to BL21 E. coli via isopropyl-beta-D-thiogalactoside (IPTG) induction to produce GST-CCL3L1 fusion protein, which was further detected by SDS-PAGE and Western blotting.
Results:
As shown and confirmed by restriction endonuclease digestion analysis, CCL3L1 was correctly inserted into pGEX-4T-1 vector. The expressed fusion protein had a relative molecular weight of approximately 34 kD.
Conclusion:
GST-CCL3L1 fusion protein can be successfully expressed using appropriate vector.
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