Structural Analysis of Human FUT4 Promoter

YANG Xue-Song, LIU Shuai, Yan-Qiu

Chinese Journal of Biochemistry and Molecular Biology ›› 2012, Vol. 28 ›› Issue (4) : 365-369.

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PDF(432 KB)
Chinese Journal of Biochemistry and Molecular Biology ›› 2012, Vol. 28 ›› Issue (4) : 365-369.
Research Papers

Structural Analysis of Human FUT4 Promoter

  • YANG Xue-Song, LIU Shuai, YAN Qiu*
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Abstract

LeY is a difucose oligosaccharide highly expressed in the most cancer in the epithelial tissues, such as breast cancer, ovarian cancer. Fucosyltransferase Ⅳ(FUT4) is the key enzyme for synthesis of LeY. Previous study showed that FUT4 improved the cell proliferation through increasing the synthesis of LeY. But the mechanism of transcription regulation of FUT4 remained unclear. In this study, we constructed plasmid with report gene containing FUT4 promoter segments with different lengths according to the bioinformatics analysis and identified their transcriptional activities in different cell lines. Sequence of FUT4 promoter was obtained according to the analysis of First EF program. Human FUT4 promoter segments in different lengths were amplified by PCR. The products of PCR were cloned into pGL6 vector after double digestion by KpnⅠ and NheⅠ, and the recombinant plasmid of different promoter length were obtained, respectively. The recombinant plasmid was transfected into MCF-7 cells and MDA-MB-231 cells with lipofectamine 2 000.The transcriptional activities were analyzed by double luciferase method. The luciferase activities of pGL6-FUT4-1.2 kb was increased in both cell lines(P<0.05). The sequence -800~-1 600 bp may be responsible for the transcriptional activities of FUT4.

Key words

fucosyltransferase Ⅳ / transcriptional regulation / promoter / reporter gene

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YANG Xue-Song, LIU Shuai, Yan-Qiu. Structural Analysis of Human FUT4 Promoter[J]. Chinese Journal of Biochemistry and Molecular Biology, 2012, 28(4): 365-369

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Funding

Supported by National Natural Science Foundation of China(No.30800195, No.31070729)

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