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Dairy-derived antihypertensive 7 peptide tandem gene

A tandem gene, anti-hypertensive technology, applied in the field of genetic engineering

Inactive Publication Date: 2011-01-19
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, so far, all milk-derived antihypertensive small peptides are obtained by protease hydrolysis of milk protein, and there is no research report on the expression of antihypertensive small peptides by genetic engineering technology

Method used

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  • Dairy-derived antihypertensive 7 peptide tandem gene
  • Dairy-derived antihypertensive 7 peptide tandem gene
  • Dairy-derived antihypertensive 7 peptide tandem gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0061] 1). Design of milk-derived antihypertensive 7-peptide tandem gene suitable for expression in Pichia pastoris

[0062] According to the amino acid sequence of the milk-derived antihypertensive 7-peptide AVPYPQR, Pichia pastoris codon usage and high-expression superior codons, the milk-derived antihypertensive 7-peptide tandem gene is designed, and there is no gap between the nucleotide sequences encoding the 7-peptide to ensure The expression product can be digested with trypsin to form a single milk-derived antihypertensive 7-peptide with complete N-terminal and C-terminal. In order to be suitable for the secretory expression of the gene in Pichia pastoris, a yeast secretory signal peptide cleavage site was introduced at the 5' end of the tandem gene. In order to facilitate the construction of the expression vector, XhoI and NotI restriction sites were respectively introduced at both ends of the gene. That is, the structure of the designed tandem gene is: XhoI-signal c...

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Abstract

The invention provides a milk antihypertensive 7 peptide series gene suitable to be expressed in a Pichia pastoris and a synthesizing method thereof. The structure of the series gene is as follows: XhoI-signal cutting sequence-7 peptide amino acid sequence repetition-stop codon-NotI; a target gene sequence is CTC GAG AAAAGA GAG GCT GAA GCT GCT GTT CCA TAC CCA CAAAGA(GCT GTC CCA TAC CCA CAA AGA GCT GTT CCA TAC CCA CAA AGA)n TAA GCGGCCGC; wherein, n refers the sequences in the bracket to be repeated for n times. The synthesizing method is as follows: 1) the designing of the milk antihypertensive 7 peptide series gene suitable to be expressed in the Pichia pastoris; and 2) the synthesizing of the milk antihypertensive 7 peptide series gene. The gene can be expressed into the Pichia pastoris in a high level; the expression product thereof after the restriction enzyme of a trypsinase can form the single milk antihypertensive 7 peptide with both the N end and the C end being complete.

Description

(1) Technical field [0001] The invention relates to a genetic engineering technology, in particular to a milk-derived antihypertensive 7-peptide tandem gene designed and synthesized by genetic engineering means, the gene can be expressed at a high level in Pichia pastoris, and the expressed product is digested with trypsin After that, a single milk-derived antihypertensive 7-peptide with complete N-terminal and C-terminal can be formed. (2) Background technology [0002] Hypertension is a chronic cardiovascular and cerebrovascular disease and a major risk factor for coronary heart disease, stroke and other cardiovascular and cerebrovascular diseases. According to statistics, the number of deaths from cardiovascular and cerebrovascular diseases caused by hypertension exceeds 12 million every year in the world. Hypertension has become the number one killer of human health and a major global public health problem. At present, the first-line drugs approved by the World Health ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12A61P9/12C07K7/06C12N15/81C12N15/10
Inventor 李杰韩建春滕春红路博李名远
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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