Overview[1]
The inorganic pyrophosphatase (PPA) gene is a gene that regulates sucrose synthesis in the process of plant sugar metabolism. In order to apply this gene to the study of sugarcane transgenics, this gene was cloned from baker’s yeast. Inorganic pyrophosphatase (yeast) was purified from a recombinant E. coli strain carrying a fusion gene of the ppa gene cloned from Saccharomyces cerevisiae and the Mycobacterium xenopi GyrA intein. This pyrophosphatase catalyzes the hydrolysis of inorganic pyrophosphate to form orthophosphate: P207-4 + H20 → 2HP04-2. In nucleic acid amplification experiments, it can relieve the inhibitory effect of the generated inorganic pyrophosphate on the reaction system. Developed by Biohelix Corporation (now a wholly owned subsidiary of Quidel Corporation). Inorganic Pyrophosphatase (English: Inorganic Pyrophosphatase, referred to as pyrophosphatase) is an enzyme that catalyzes the conversion of one molecule of pyrophosphate into two molecules of phosphate ions. This is a highly exergetic reaction, so this reaction can be coupled to some thermodynamically unfavorable transformations in order to drive these transformations to completion. The function of this enzyme is to play an important role in lipid metabolism (including lipid synthesis and breakdown), calcium absorption, bone formation and DNA synthesis, as well as other biochemical transformations.
Components
▼▲
Cabot Carbon Black
Component name
Quantity
Yeast Pyrophosphatase (0.1 U/μl)
100 μl/1 ml
Application
Improving RNA yield in reverse transcription reactions.
Enhanced DNA replication.
Activity definition
1 unit refers to the amount of enzyme required to catalyze inorganic pyrophosphate to produce 1 μmol of phosphate per minute under standard reaction conditions (20mM Tris-HCl, pH 7.5, 2mM MgCl2, 2mM PPi, 25°C for 10 minutes).
Save
Can be stored at -20℃ for 3 years.
Precautions for use
(1) This enzyme is active in a variety of reaction buffers. Usually, this enzyme can be directly connected in experiments such as HDA amplification and LAMP amplification.
(2) The dosage of this enzyme needs to be optimized in different experiments, usually at a concentration of 0.05~1U/ml.
(3) The optimal reaction temperature of this enzyme is 25°C. It is active at 16~37°C. The enzyme can be inactivated at 65°C for 10 minutes.
Main references
[1] Fan Haikuo, Huang Dongjie, Liu Qiaoquan, et al. Cloning analysis of inorganic pyrophosphorylase fusion gene and construction of expression vector [J]. Journal of Southwest Agriculture, 2007, 20(6): 1267-1271.
