Method for constructing Exendin-4 high-yielding engineering bacterial strain

Abstract

The invention relates to the alteration of an Exendin-4 target gene and the construction of an expression vector, belonging to the field of genetic engineering. The method comprises the following steps: alternating 70 percent of codons of an original gene according to the amino acid sequence of the Exendin-4 in the gene bank and combining with the preference ranking of codons of colon bacillus so that the gene can be efficiently expressed in the colon bacillus; adding proper enzyme cutting sites and enterokinase sites to establish the proper expression vector; converting a recombined expression vector into different colon bacillus engineering bacteria; and selecting bacterial strains capable of efficiently expressing the Exendin-4.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a method for constructing an Exendin-4 high-yielding engineering strain. Background technique [0002] Diabetes is a worldwide disease. There are more than 150 million diabetic patients in the world, of which 90% are type 2 diabetic patients. There are nearly 40 million type 2 diabetes patients in our country, and with the improvement of living standards, the incidence rate will become higher and higher. Currently, the drugs (western medicines) for treating type 2 diabetes mainly include sulfonyls, biguanides, α-glucosidase inhibitors, glitazone insulin sensitizers, and glinide insulin secretagogues. These drugs have different degrees of side effects, such as insulin secretion-stimulating drugs, which promote the secretion of insulin by the pancreas and do not depend on the level of blood sugar concentration, not only produce symptoms of fasting hypoglycemia, but also cause pa...

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Problems solved by technology

People such as Xiaopu Yin of East China University of Science and Technology once transformed the Exendin-4 gene into Escherichia coli, but the yield was extremely low, and finally the recombinant Exendin-4 with a purity of 80% was only 3.14mg / Kg bacteria (Xiaopu Yin, Dongzhi Wei, Lina Yi, Expression and purification of Exendin-4, a GLP-1 receptor agonist, in Escherichis coli, Protein Expression and Purification, 2005, 41: 259-265.)

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