转录因子STAT5A启动子荧光素酶报告基因质粒的构建及鉴定

doi:10.3969/j.issn.1004-616x.2022.01.010

Abstract

Abstract: OBJECTIVE: To construct and to evaluate activities of luciferase reporter plasmids for transcription factor STAT5A promoter. METHODS: The sequence for the promoter region in STAT5A gene was amplified by PCR using genomic DNA of the breast cancer cell line MCF7 as a template and cloned into luciferase reporter plasmid pGL3-Enhancer. The target plasmid was amplified by bacterial fluid,examined by enzyme digestion and direct sequencing. The function of target plasmid was further verified by dual luciferase reporter gene assay. RESULTS: It was found that the cloned sequence of STAT5A promoter region in target plasmid was correct,and the target plasmid had transcriptional activities. Dual-luciferase assays found that the relative activities of the recombinant vector pGL3-STAT5A-pro-luc-E luciferase were about 8 times that of the negative control pGL3-Enhancer (P<0.01),and the pGL3-STAT5A-pro-N-luc-E luciferase activities were about 6 times that of the negative control pGL3-Enhancer (P<0.01). CONCLUSION: A luciferase reporter plasmid for transcription factor STAT5A promoter was successfully constructed. The plasmids would provide an important research tool for further study of STAT signal transduction and the transcriptional activator protein signaling pathway.

Key words: STAT5A, promoter, reporter gene, luciferase, PCR

CLC Number:

Q783

CHEN Minna, LI Chunlan, LIU Jing. Construction and identification of luciferase reporter plasmids for the transcription factor STAT5A promoter[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2022, 34(1): 53-56,71.

References

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Construction and identification of luciferase reporter plasmids for the transcription factor STAT5A promoter

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