[Expression of (His)(6)-eIF3s4 fusion protein in human breast cancer cell Bcap37].

Abstract

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.

• Authors: Qun Wei, Feng-jia Zhu, Yi Wang, Ping Chen, Lin-bo Wang, Jiang Cao
• Journal: Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology (Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi) Vol. 23 Issue 7 Pg. 627-9 (Jul 2007) ISSN: 1007-8738 [Print] China
• PMID: 17618584 (Publication Type: Journal Article)

Chemical References

• DNA, Complementary
• Eukaryotic Initiation Factors
• PHB2 protein, human
• Prohibitins
• Recombinant Fusion Proteins

Topics

• Blotting, Western
• Breast Neoplasms
• Cell Line, Tumor
• Cloning, Molecular
• DNA, Complementary (genetics)
• Eukaryotic Initiation Factors (genetics, metabolism)
• Genetic Vectors (genetics)
• Humans
• Prohibitins
• Recombinant Fusion Proteins (genetics, metabolism)
• Reverse Transcriptase Polymerase Chain Reaction