Human parathyroid hormone eukaryotic expression recombinant plasmid vector and construction method thereof

Abstract

The invention relates to the field of gene medicines, and particularly provides a human parathyroid hormone eukaryotic expression recombinant plasmid vector and a construction method thereof. The construction method comprises the following steps of artificially synthesizing a parathyroid hormone (hPTH1-84aa) gene (SEQ ID NO.1), carrying out PCR amplification to obtain parathyroid hormone gene segments, carrying out double enzyme digestion on a PCR product by using restriction enzymes BamHI and XhoI, connecting the product with an eukaryotic expression vector plasmid PHY-810 subjected to the same enzyme digestion, converting the connected expression vector into competent bacteria, and carrying out coating and screening to obtain the human PTH gene eukaryotic expression recombinant plasmid vector (figure 1: PHY-PTH). The eukaryotic expression recombinant plasmid vector constructed by the method enables cells to highly express PTH, and can be used for gene therapy of hypoparathyroidism and osteoporosis.

Description

technical field [0001] The invention relates to the field of gene medicine, in particular to the construction of a recombinant plasmid vector for gene therapy of hypoparathyroidism. Background technique [0002] Parathyroid hormone is an active polypeptide hormone secreted by the human parathyroid gland. Its main physiological function is to maintain blood calcium balance by regulating bone cells and kidney cells, and stimulate kidney reabsorption of calcium, phosphorus secretion and bone reconstruction. It is an important peptide hormone that regulates calcium and phosphorus metabolism. [0003] Hypoparathyroidism is a disorder of calcium and phosphorus metabolism caused by insufficient secretion of parathyroid hormone caused by disease, aging and surgical injury. Hypoparathyroidism caused by disease and aging is characterized by decalcification, bone regeneration disorder, and osteoporosis; hypoparathyroidism caused by surgical injury is divided into temporary type and pe...

AI Technical Summary

Problems solved by technology

[0006] However, the existing recombinant human parathyroid hormone preparations have deficiencies, which affect clinical use

First of all, recombinant PTH metabolizes quickly in the body and is very unstable. It needs regular daily medication to maintain blood drug concentration, which increases the difficulty of medication and affects the curative effect. , nausea and vomiting; the greater limitation is that the treatment of this product cannot exceed 24 months at the longest, and the patient can only be introduced once in a lifetime for a maximum of 24 months of treatment

The third is the high cost of preparation: due to the low content of PTH expression products, separation and purification are difficult, resulting in high preparation costs and expensive drugs, which limit large-scale use

No patents have been found on the construction of PTH recombinant plasmid expression vectors for gene therapy through patent searches

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