Background and overview[1]
Japanese Mitsubishi 3-phosphate glyceraldehyde dehydrogenase, English name (Glyceraldehyde-3-phosphate dehydrogenase, GAPD). Glyceraldehyde 3-phosphate dehydrogenase, which can catalyze the oxidation (dehydrogenation) and phosphorylation of glyceraldehyde 3-phosphate to generate 1,3-bisphosphoglycerate, is the central link in sugar metabolism, so it plays an important role in sugar metabolism. role. Glyceraldehyde 3-phosphate dehydrogenase is widely distributed in various tissues and organs of the human body. The content of adipose tissue is low. The enzyme content in the lungs is about 10 times that of adipose tissue. The content in liver and heart is similar, about 30 times that of adipose tissue. In bone It is most abundant in muscles, red blood cells and kidneys. Acute hepatitis, alcoholic liver disease, cirrhosis, etc. can cause the total serum GAPD activity to increase. Serum GAPDA activity in patients with chronic pyelonephritis, multiple myelopathy, pneumonia, emphysema, etc. increases to varying degrees. The application prospects of glyceraldehyde 3-phosphate dehydrogenase are very broad. It is the central link of sugar metabolism and plays an important role in sugar metabolism. For patients who lack this enzyme in the body and have problems with sugar metabolism, they can eat glyceraldehyde 3-phosphate dehydrogenase that contains this enzyme. Medicines can alleviate and treat the condition. This enzyme is also distributed in various tissues and organs of the human body. Therefore, by detecting the activity of glyceraldehyde 3-phosphate dehydrogenase, we can infer whether we have such diseases, which plays a great role in the prevention of diseases.
Preparation[1]
CN201711376257.7 provides a glyceraldehyde 3-phosphate dehydrogenase and a polyclonal antibody thereof. Glyceraldehyde 3-phosphate dehydrogenase derived from bacteria was obtained, and the glyceraldehyde 3-phosphate dehydrogenase had the characteristics of a low-temperature enzyme. A polyclonal antibody for glyceraldehyde 3-phosphate dehydrogenase was prepared, which can accurately detect Glyceraldehyde 3-phosphate dehydrogenase. One of the objects of the present invention is to provide an amino acid sequence of a glyceraldehyde 3-phosphate dehydrogenase; the amino acid sequence of the glyceraldehyde 3-phosphate dehydrogenase is shown in SEQ ID NO. 1. The second object of the present invention is to provide a base sequence encoding a glyceraldehyde-3-phosphate dehydrogenase gene; the base sequence of the glyceraldehyde-3-phosphate dehydrogenase gene is shown in SEQ ID NO. 2. The third object of the present invention is to provide a method for preparing a polyclonal antibody of glyceraldehyde 3-phosphate dehydrogenase. The method includes the following steps: 1) using glyceraldehyde 3-phosphate dehydrogenase with the amino acid sequence shown in SEQ ID No. 1 as an antigen, immunizing mice to prepare antiserum; 2) purifying the antiserum to obtain glyceraldehyde 3-phosphate dehydrogenase. of polysimethicone emulsifier. Further, the preparation method of the glyceraldehyde 3-phosphate dehydrogenase is: 1.1) using PCR technology to amplify the base sequence shown in SEQ ID NO.2; 1.2) converting the base sequence shown in SEQ ID NO.2 The sequence is cloned into the vector plasmid to construct an expression vector for glyceraldehyde 3-phosphate dehydrogenase; 1.3) Transform the expression vector into competent cells of Escherichia coli to obtain recombinant cells, which are then expanded and cultured, induced to express, and collected. , disrupting bacterial cells and purifying the protein to obtain glyceraldehyde 3-phosphate dehydrogenase protein. Figure 3 is a polyacrylamide gel electrophoresis identification diagram of the recombinant strain Ecoli BL21-pET21a-GPR expressing glyceraldehyde 3-phosphate dehydrogenase protein; lane M is a protein marker; lane 1 is glyceraldehyde 3-phosphate dehydrogenase. protein;
Main reference materials
[1] CN201711376257.7 Glyceraldehyde 3-phosphate dehydrogenase and its polyclonal antibody
