System and method for detecting overexpression reconstructed gene plasmid in expression and phagocyte

• Main Authors: GE XIANG, BU GUOYUN, HAN XIAOLEI, XU HAIWEI, HUO ZHENXIN, CHEN CHEN, ZHANG XI, LI TONGTONG, BAI SHUCAI
• Format: Patent
• Language: Chinese; English
• Published: 12.01.2024
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; VINEGAR; WINE

The invention belongs to the technical field of gene engineering detection, and discloses a system and a method for detecting overexpression reconstructed gene plasmids in expressed and phagocytic cells. The method comprises the following steps: extracting nucleic acid samples from expression cell lines and phagocytes; carrying out PCR (Polymerase Chain Reaction) amplification by using the existing specific primer; fluorescent real-time quantitative PCR is realized by using an existing fluorescent probe; and determining the expression level of the circ0058514 or the intake of the circ0058514 in the phagocytes according to the intensity and change trend of a fluorescence signal in a PCR reaction system. Conventional laboratory equipment and reagents are used, operation is easy, and the method is suitable for large-scale laboratory research and clinical application. 本发明属于基因工程检测技术领域，公开了过表达重构基因质粒在表达、吞噬细胞中检测系统及方法。该方法包括：从表达细胞系和吞噬细胞中提取核酸样本；使用现有特异性引物进行PCR扩增；使用现有荧光探针实现荧光实时定量PCR；根据PCR反应体系中荧光信号的强度和变化趋势测定circ_0058514的表达水