Recombinant human pepsinogen II isozyme chimeric protein, and preparation method and applications thereof

Abstract

The invention discloses a recombinant human pepsinogen II isozyme chimeric protein, and a preparation method and applications thereof. The preparation method comprises the steps as follows: by taking gene sequences of two isozymes of recombinant human pepsinogen II as a template, carrying out PCR (polymerase chain reaction) splicing to obtain a chimeric protein coding gene sequence, constructing recombinant expression plasmids, transforming the screened positive clone plasmids into expression host cells, screening an efficiently-expressed recombinant chimeric protein strain, carrying out enlargement culture on the cells and inducing expression chimeric protein, and purifying to obtain the recombinant human pepsinogen II isozyme chimeric protein. The invention further discloses the applications of the recombinant human pepsinogen II isozyme chimeric protein in preparing monoclonal antibodies and multiresistant serums or pepsinogen II kit calibration products. A great amount of stably expressed recombinant human pepsinogen II isozyme chimeric protein can be produced by utilizing a genetic engineering technology, and one protein has two chimeric protein sequences of the human pepsinogen II.

Description

technical field

[0001] The invention belongs to the field of biotechnology, and specifically relates to a recombinant human pepsinogen II isozyme chimeric protein, a preparation method and an application thereof. Background technique

[0002] Pepsinogen (PG) is a proteolytic zymogen secreted by gastric mucosal cells, which belongs to the aspartic proteinase family. Under acidic conditions with a pH of 1.0-5.0, it can be automatically hydrolyzed into pepsin. According to the biochemical and immunological characteristics, it can be divided into two categories: PG I (PGA) and PG II (PGC). There are 7 groups of pepsin isoenzymes. Secreted by the mucus neck cells of the principal cells of the gastric glands; components 6 to 7 are called PG II, derived from the oxyntic glands of the gastric fundus mucosa of the gastric body, gastric cardia glands, gastric antrum pyloric glands and distal duodenal Brunner's glands, The prostate and pancreas also produce small amounts. Most of th...

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