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Adiponectin-Glucagon-like peptide-1-like peptide recombinant protein expression vector and construction

A technology of adiponectin and fusion gene, which is applied in the biological field, can solve the problems that the half-life in vivo has not been improved accordingly, the mechanism of GLP-1 clearance cannot be explained, and the half-life in vitro has been prolonged, so as to achieve the effect of maintaining spatial conformation and biological activity

Inactive Publication Date: 2006-02-22
ZHEJIANG UNIV
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Problems solved by technology

However, only the participation of dipeptide peptidase IV cannot explain all the clearance mechanisms of GLP-1 in vivo. The half-life of GLP-1 in plasma in vitro is 20 minutes, far exceeding the in vivo half-life of 3 to 11 minutes obtained by many studies.
The in vitro half-life of GLP-1-A obtained by Deacon et al. by substituting glycine for the eighth amino acid alanine of GLP-1 (7-37) was significantly prolonged, but the in vivo half-life was not correspondingly improved, suggesting that there are other mechanisms Involved in the clearance of GLP-1 in vivo

Method used

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  • Adiponectin-Glucagon-like peptide-1-like peptide recombinant protein expression vector and construction
  • Adiponectin-Glucagon-like peptide-1-like peptide recombinant protein expression vector and construction
  • Adiponectin-Glucagon-like peptide-1-like peptide recombinant protein expression vector and construction

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Embodiment 1

[0032] 1. Materials and Instruments

[0033] 1.1 Materials

[0034] Escherichia coli strain E.coli DH5α, BL21(DE3) was provided by the Cancer Institute of the Second Affiliated Hospital of Zhejiang University, the prokaryotic expression plasmid PET28a was purchased from Novagen, Germany, the RNA extract (Trizol) was purchased from Gibcol BRL, USA, and pyrocarbonic acid Diethyl ester, Oligo(dT) 15 , UNIQ-10 Column Genomic DNA Extraction Kit were purchased from Shanghai Sangon Bioengineering Company, M-MLV Reverse Transcriptase, RNasin, dNTPs, Taq DNA polymerase, agarose, T 4DNA ligase, restriction endonuclease NheI, BamHI, HindIII, XbaI were purchased from Promega Company of the United States, lymphocyte extract was purchased from Shanghai Hengxin Chemical Reagent Co., Ltd., GeneRuler TM DNA Ladder Mix was purchased from Fermentas Company in Germany, DNA Marker DL2000, Isopropyl-beta-D-thiogalactopyranoside (IPTG) was purchased from TaKaRa Company in Japan, SDS polyacrylamid...

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Abstract

Disclosed is an adiponectin-Glucagon-like peptide-1-like peptide recombinant protein expression vector and construction, wherein adiponectin and GLP-1-A encoded gene are obtained by utilizing PCR technology, the method comprises obtaining adiponectin through amplification of adiponectin spherical domain gene, then obtaining GLP-1-A encoded gene by mutating codons encoding No. 8 amino acid alanine in GLP-1-(7-37) encoded gene, then cloning GLP-1-A and adiponectin encoded genes directionally to expression vector by utilizing enzyme digestion technology, obtaining adiponectin-glucagon-like peptide-1 analogue peptide fusion protein expression vector, finally expressing adiponectin-GLP-1-A fusion protein through transforming protein expression bacteria.

Description

technical field [0001] The invention relates to the field of biology, in particular to adiponectin-glucagon-like peptide-1 similar peptide fusion protein expression vector and construction method. Background technique [0002] Type 2 diabetes (Diabetes Mellitus, DM) is a disease that seriously endangers human health. The number of patients is increasing rapidly with the improvement of living standards, the aging of the population, and the advancement of diagnostic techniques. Recent studies have shown that insulin resistance and insulin insufficiency are the characteristic pathophysiological changes of type 2 diabetes, and the interaction between the two eventually leads to the occurrence of type 2 diabetes. Not only that, insulin resistance is also involved in the occurrence and development of chronic complications of diabetes, such as cardiovascular complications, diabetic nephropathy, etc., and is an important hub linking diabetes, hypertension, hyperlipidemia, and corona...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C07K19/00C12N15/70C12N15/74
Inventor 赵同峰谷卫詹宇红赵江佩
Owner ZHEJIANG UNIV
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