Method for producing gene engineering recombination 192 peptide human growth hormone

Abstract

The invention belongs to the technical field of molecular biology, in particular to a method for producing genetic engineering recombinant 192 peptide human growth hormone. The technical scheme is as follows: (1) Obtaining the 192-peptide human growth hormone gene: using the pHil-D2 plasmid as a shuttle plasmid, the growth hormone cDNA is transferred into yeast cells, and through homologous recombination with genes on the chromosomes of yeast cells, The genetically engineered hGH yeast was constructed; (2) expression system: the strain of Pichia pastoris was used as the expression host to express 192 peptide human growth hormone in the way of intracellular expression; (3) the hGH yeast was induced to express and produce human growth hormone.

Description

Technical field: [0001] The invention belongs to the technical field of molecular biology, in particular to a method for producing genetic engineering recombinant 192 peptide human growth hormone. Background technique: [0002] Human growth hormone (hGH) is a protein hormone secreted by eosinophilic cells in the anterior pituitary gland, consisting of 191 amino acids. Normal adult plasma hGH level is about 1~5ng / ml, half-life is 15~20 minutes. [0003] hGH is one of the hormones with the highest content in the human body. Because it affects the growth of cells, bones, muscles and organs throughout the body, it has a wide range of regulatory effects on human metabolism and various physiological functions. For example: hGH promotes fat decomposition and makes blood plasma Elevation of free fatty acids; promote the synthesis of RNA and protein in many tissues, a positive nitrogen balance occurs, and make cartilage, bone, muscle connective tissue and viscera and other tissues s...

AI Technical Summary

Problems solved by technology

The hGH expressed in Escherichia coli forms inclusion bodies and becomes insoluble, which is difficult to extract and requires complex steps of denaturation and renaturation, so the loss is very large; some are secreted expression, but the expression host of Escherichia coli contains endotoxin, in order to remove Removal of endotoxin requires more complicated purification steps, resulting in a lower product yield, and the crude product still contains a large amount of E. coli impurity proteins, especially the expression host-specific E. coli endotoxin

In order to remove these components harmful to the human body, it is necessary to go through complicated purification steps, the loss is also very large, and the final product yield is very low (25mg / L)

High production costs due to inherent flaws in this expression system