Hypoxia regulation of the report and identification of vector.docVIP

 PAGE \* MERGEFORMAT 10 Hypoxia regulation of the report and identification of vector Authors: Yang, Li Zhanting, Alex, Chai Qing-huan, Wang, Zhang Hui, Sun Ji-Feng [Keywords:] luciferase Construction and identification of hypoxia regulatory reporter vectors [Abstract] AIM: To construct the firefly luciferase reporter gene vectors regulated by hypoxia response promoters. METHODS: The oligonucleotides of hypoxia response element (HRE) and a mutation control were synthesized and cloned into pCIneo vectors to constrct HRE / CMV recombinant vectors. Then HRE / CMV promoters amplified by PCR were subcloned into pGL3Basic vectors. RESULTS: The integrity reporter vectors were verified by restriction enzyme digestion. Also, the sequences of the vectors were detected and aligned to the expected sequences. CONCLUSION: Luciferase reporter vectors regulated by HRE / CMV promoters were successfully constructed, which makes it possible to further investigate their potency of hypoxia regulation. [Keywords] transcription; regulation; hypoxia; response elements; luciferase; reporter gene [Abstract] Objective: To construct can be used to detect hypoxia response promoter transcriptional regulation role of the firefly luciferase reporter vector. Methods: The synthesis of hypoxia-responsive element (HRE) oligonucleotide sequence and its mutant control, and cloned into pCIneo , build HRE / CMV recombinant vector. and then use PCR method HRE / CMV promoter subcloned into the pGL3Basic. Results: The restriction enzyme digestion and sequencing analysis, the cloned gene product in line with expectations digested the results, serial no base mutations. Conclusion: The successful construction of the hypoxia response promoter luciferase reporter vector, in order to further study its role in providing the conditions for the hypoxic control. [Keywords:] transcription; control; hypoxia; response element; luciferase; reporter gene 0 Introduction Has developed a variety of reporter vect