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Construction method of recombinant human albumin and expression vector thereof

A technology of human serum albumin and expression vectors, which is applied in the field of construction of recombinant human serum albumin and its expression vectors, and can solve problems such as the contradiction between supply and demand, and the shortage of blood product supply

Pending Publication Date: 2016-12-14
NORTHWEST UNIVERSITY FOR NATIONALITIES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the influence of many factors such as the restructuring of the plasma station and the strengthening of the safety supervision of blood products, the supply of blood products represented by human serum albumin is seriously in short supply in the domestic market, and the contradiction between supply and demand is prominent.

Method used

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  • Construction method of recombinant human albumin and expression vector thereof
  • Construction method of recombinant human albumin and expression vector thereof
  • Construction method of recombinant human albumin and expression vector thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1. Construction of human albumin mammalian eukaryotic expression vector

[0031] 1. Acquisition of human serum albumin (HSA) gene

[0032] Inquire about HSA-related information and literature on GENBANK, GOOGLE Scholar, etc., HSA gene accession number: NM_000477.5.

[0033] Preproalbumin has a total of 609 amino acids, and the accession number is NP_000468.1, of which the first 24 encode signal peptides (short peptide chains (5-30 amino acids in length) that guide the transfer of newly synthesized proteins to the secretory pathway.

[0034] 2. Construction of recombinant expression vector pcDNA3.1-HSA

[0035] According to the codon preference of mammalian cells, mainly codon optimization for CHO cells, upstream insertion Kpn I restriction site, downstream insertion xho I restriction site, and in Kpn The Kozak sequence was added after the restriction site I, and the target gene was synthesized by Shanghai Sangon Biotechnology Co., Ltd. with a full length of 1845...

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Abstract

The invention provides a recombinant human albumin with an amino acid sequence of SEQIDNo.2 in the sequence table. The invention also provides a construction method of the recombinant human albumin expression vector. The recombinant human albumin expression vector prepared by the method can secrete human albumin. The expression vector is optimized, and the expression of HSA reaches 150-180 mg / L.

Description

technical field [0001] The invention relates to the construction method of recombinant human serum albumin and its expression vector. Background technique [0002] Human serum albumin (HSA) is the most abundant protein in human plasma, accounting for 40%-60% of total plasma protein, and plays an important role in maintaining plasma colloid osmotic pressure and transporting nutrients in the body. Clinically, human albumin is mainly used for blood loss trauma, shock caused by burns, cerebral edema and increased intracranial pressure caused by injury, edema or ascites caused by liver cirrhosis and nephropathy; in cardiopulmonary bypass, adjuvant treatment of burns, hemodialysis It is also widely used in adjuvant therapy and adult respiratory distress syndrome. It is an essential drug for hospital clinical emergency or severe surgery, and has always occupied a pivotal position in the hospital market. Human albumin is usually extracted from plasma. Due to the influence of many ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765C12N15/70C12N15/85C12N15/66
CPCC07K14/765C12N15/66C12N15/70C12N15/85C12N2800/101C12N2800/107
Inventor 马忠仁李向茸冯若飞张海霞李明生乔自林包晓婧马玉梅令瑛
Owner NORTHWEST UNIVERSITY FOR NATIONALITIES