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Method for identifying transcriptional activity of EFTUD2 promoter

An identification method and promoter technology, applied in biochemical equipment and methods, microbial measurement/testing, recombinant DNA technology, etc., can solve problems such as no research reports

Active Publication Date: 2019-10-18
江苏纳华生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, it is of great biological significance to study and elucidate the expression regulation mechanism of EFTUD2 gene, but there is no relevant research report so far.

Method used

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  • Method for identifying transcriptional activity of EFTUD2 promoter
  • Method for identifying transcriptional activity of EFTUD2 promoter
  • Method for identifying transcriptional activity of EFTUD2 promoter

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Experimental program
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Embodiment 1

[0078] 1. Main experimental materials

[0079] Human hepatoma cell HepG2 is preserved in the applicant's laboratory, and conventional culture uses DMEM medium (Gibco Company, the United States) containing 10% fetal bovine serum (Gibco Company, the United States), and the medium has added 1.5mM L-Glutamine, 100U / mlpenicillin, 100 μg / ml Streptomycin (sigma), at 37°C, 5% CO 2 cultured in a saturated humidity incubator.

[0080] 10cm culture dishes and 96-well plates were purchased from Corning; PBS was purchased from amerosco; Trypsin-EDTASolution (0.25% Trypsin+0.53mM EDTA, Hyclone); GP-transfect-Mate transfection reagent, plasmid pUC57 and psiCHECK-2 were all purchased from Genepharma; DNA endonuclease, DNA ligase and DNA marker were purchased from Fermentas; agarose and DNA gel recovery kits were purchased from Tiangen Biochemical Technology Co., Ltd.; medium extraction kits were purchased from Hangzhou Ai Sijin Biotechnology Co., Ltd.; yeast extract and tryptone were purch...

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Abstract

The invention provides a method for identifying transcriptional activity of an EFTUD2 promoter. The method comprises the following steps of (1) construction of an EFTUD2 promoter reporter gene recombinant vector, (2) transient transfection and (3) detection of dual luciferase reporter gene activity. The method for identifying the transcriptional activity of the EFTUD2 promoter lays a solid foundation for elucidating the expression regulation mechanism of an EFTUD2 gene, and also is conductive to further studying the relationship between the expression level of the EFTUD2 and innate immunity and a specific mechanism of differential splicing.

Description

technical field [0001] The invention relates to the technical field of biological genes, in particular to a method for identifying the transcriptional activity of an EFTUD2 promoter. Background technique [0002] The EFTUD2 (Elongation factor Tu GTP-binding domain-containing2) gene encodes a GTPase, which is a component of the U5 small nuclear ribonucleoprotein (snRNP), the spliceosome complex, together with the rest of the spliceosome Play a role in splicing precursor mRNA (precursormessager RNA, pre-mRNA) to produce mature mRNA. During the splicing process, EFTUD2 can be directly involved in the splicing process as well as in the spliceosomal snRNP recycling process. As a kind of GTP hydrolase, EFTUD2 acts like a spliceosome translocase, which can not only mediate the conformational change of RNA or protein, but also promote the base pairing of U1 and the 5' splice site of the pre-RNA, and promote U4 and U6 Base pairing is separated to form a 5' splice site, U6 and U2 ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C12N15/66C12Q1/686C12Q1/66
CPCC12N9/14C12N15/66C12N15/85C12Q1/66C12Q1/686C12Y306/05003
Inventor 朱传龙李毓雯徐瑞瑞宁琴罗小平李军胡平平
Owner 江苏纳华生物科技有限公司
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