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 sothat 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

Construction method of Exendin-4 high-yielding engineering strain technical field 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 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...

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Problems solved by technology

People such as XiaopuYin 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 (XiaopuYin, DongzhiWei, LinaYi, ExpressionandpurificationofExendin-4, aGLP-1 receptoragonist, in Escherichiscoli, Protein Expression and Purification, 2005, 41: 259-265.)

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