Method for preparing alanine aminotransferase and application

Abstract

The invention discloses a method for preparing alanine aminotransferase, which includes the following steps: modifying a whole-genome-synthesized humanized alanine aminotransferase encoding gene sequence, and removing rare codons of escherichia coli; inserting the modified alanine aminotransferase encoding gene sequence into an expression vector, and adopting bacterial strains of escherichia coli for transformation and expression; inoculating the transformed and expressed bacterial strains with an LB culture medium and performing IPTG (isopropyl-Beta-d-thiogalactoside) induction; and breaking the obtained thalli, performing centrifugal separation, and passing by the Ni-NTA columns for purification. The operation for preparing alanine aminotransferase is very simple, the preparation cost is low, the product yield is high, the purity is high, and high biological activity is obtained; the prepared alanine aminotransferase serving as antigenic substances can be used for animal immunization, related antibodies can be screened; and meanwhile, the method lays the foundation of quick establishment for a calibrator preparing platform imperative to the domestic clinical chemistry diagnosis as well as development and preparation for diagnostic reagent standard substances and references.

Description

technical field [0001] The invention belongs to the field of genetic engineering and protein expression, and relates to a method for efficiently preparing alanine aminotransferase by using bioengineering technology. The product can be used as an antigenic substance for animal immunization, to screen related antibodies, and at the same time as a calibrator and standard for clinical chemical diagnosis. Lays the groundwork for the preparation and development of substances and reference materials. Background technique [0002] Alanine aminotransferase (alanine aminotransferase, ALT, EC2.6.1.2) is a transaminase between glutamic acid and pyruvate, also known as alanine aminotransferase, referred to as GPT and ALT. Elevated alanine aminotransferase is a common clinical phenomenon. The liver is the largest detoxification organ in the human body. Whether this organ is normal or not is very important to the human body. Elevated GPT is an important indicator of liver function proble...

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Problems solved by technology

[0007] 3. Alanine aminotransferase will also increase in heavy or long-term drinkers

Almost none of the major domestic manufacturers of diagnostic reagents has the experience and strength to produce corresponding quality control products, and there are very few domestically produced products recognized in the market

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