Metridia荧光素酶检测条件的优化

doi:10.3969/j.issn.1004-616x.2016.06.005

Abstract

Abstract: OBJECTIVE: To investigate whether the pH and serum of cell culture medium would affect the Metridia luciferase (MLuc) assay or not,and to explore a modified assay method for MLuc. METHODS: A recombinant retrovirus vector which expressed MLuc was constructed and then transiently transfected into human cervical cancer cell line HeLa to produce MLuc. Then,the effects of cell culture medium (e.g. buffers combined with reagents) on bioluminescence of MLuc were assayed. RESULTS: Various pH levels of cell culture medium caused great fluctuation of the bioluminescence intensity of MLuc. In addition,serum in the cell culture medium would alter the background of MLuc. When the cell culture medium was replaced by phosphate buffered saline (PBS) supplemented with 0.02% (V/V) NP-40 (pH=7.4),the bioluminescence intensity between them was comparable. However,the background level of MLuc was decreased from 600 to 20. CONCLUSION: Using PBS supplemented with 0.02% NP-40 as an assay buffer for MLuc,the side effects from pH and serum of cell culture medium during MLuc assay would be reduce and application of MLuc in research can be improved.

Key words: luciferase, pH, serum, assay, modify

CLC Number:

Q819

GUO Zhilan, CHE Luyang, LI Jingzhe, WU Shuang, SUN Zhenxiao, LIU Changzhen. A modified assay system for Metridia luciferase[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(6): 438-442.

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A modified assay system for Metridia luciferase

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