PCR (Polymerase Chain Reaction) primer composite for quantifying transcriptional level of mice xanthine oxidase and PCR method thereof
A technology of xanthine oxidase and transcription level, applied in the field of real-time fluorescent quantitative PCR, to achieve the effect of strong primer specificity and simple method
Inactive Publication Date: 2014-06-04
HENAN UNIV OF SCI & TECH
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At present, there is no research on the quantitative method of xanthine oxidase transcription level at home and abroad
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[0028] The PCR primer combination for quantifying the transcription level of mouse xanthine oxidase in this embodiment, the nucleotide sequence is:
[0029] Forward: 5’-TATGGCCCCGAGGTAAAAAC-3’
[0030] Reverse: 5’-GGGCGGCCTGTCTTGTATGC-3’’.
[0031] The following are specific examples of PCR primer combinations and PCR methods for quantifying the transcription level of mouse xanthine oxidase to determine the effect of molybdenum on the transcription level of mouse xanthine oxidase
[0032] 1 Experimental animals
[0033] 1.1.1 Selection of experimental animals
[0034] 60 two-month-old clean ICR mice.
[0035] 1.1.2 Grouping of experimental animals
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The invention relates to a PCR (Polymerase Chain Reaction) primer composite for quantifying the transcriptional level of mice xanthine oxidase and a PCR method thereof. The nucleotide sequence of the PCR primer composite is 5'-TATGGCCCCGAGGTAAAAAC-3' in a forward direction and 5'-GGGCGGCCTGTCTTGTATGC-3'' in a reverse direction. The PCR primer composite can be obtained by carrying out a DNA synthesis technology by utilizing a chemical synthesis method well known by technical personnel in the field, and a primer sequence is on the basis of sufficiently analyzing the gene sequence of the mice xanthine oxidase so as to have no amplification signal on a non-target gene in a material. In the invention, the transcriptional level of the mice xanthine oxidase is quantified by using the PCR primer composite, and a method is simple and accurate; and the PCR primer composite is high in specificity, can not be interfered by other genes, thereby providing an effective tool for researching the functions of the mice xanthine oxidase and the influence of other factors on the mice xanthine oxidase.
Description
Technical field [0001] The invention relates to a PCR primer combination for quantifying the transcription level of xanthine oxidase in raw mice, and also relates to a real-time fluorescent quantitative PCR method using the primer, which belongs to biological detection technology. Background technique [0002] Xanthine oxidase (XO) is an important enzyme in the metabolism of nucleic acids in the body. The enzyme is widely distributed in the plasma membrane of tissues such as the human heart, lung, liver, and small intestine mucosa. XO in serum is mainly derived from liver cells. Xanthine oxidase is an enzyme that is not highly specific. It can catalyze hypoxanthine to xanthine and then uric acid, and it can also directly catalyze xanthine to uric acid. The substrate is more specific. In addition to purine derivatives as electron donors, pteridine derivatives and aldehydes (to generate carboxylic acids) can also be used as electron donors. Hydroxy compounds are formed on the surfa...
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IPC IPC(8): C12Q1/68C12N15/11
Inventor 张才汪洋王宏伟林霖尚泽松王亚垒朱重伟
Owner HENAN UNIV OF SCI & TECH