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ApNGT gene of actinobacillus pleuropneumoniae and application of ApNGT gene

A technology for actinomycetes and pleuropneumonia, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of inability to be widely used in clinical practice, limited quantity and high cost, and achieve the effect of improving the efficiency of polypeptide glucosylation

Inactive Publication Date: 2016-04-20
NANKAI UNIV
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  • Abstract
  • Description
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Problems solved by technology

[0004] For a long time, most drug glycoproteins rely on direct extraction from organisms, but because of the limited quantity, high cost and problems such as virus infection, they cannot be widely used in clinical practice

Method used

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  • ApNGT gene of actinobacillus pleuropneumoniae and application of ApNGT gene
  • ApNGT gene of actinobacillus pleuropneumoniae and application of ApNGT gene
  • ApNGT gene of actinobacillus pleuropneumoniae and application of ApNGT gene

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Embodiment Construction

[0032] One. Describe the present invention below by specific embodiment. Unless otherwise specified, the technical means used in the present invention are methods known to those skilled in the art. In addition, the embodiments should be considered as illustrative rather than limiting the scope of the invention, the spirit and scope of which is defined only by the claims. For those skilled in the art, on the premise of not departing from the spirit and scope of the present invention, various changes or modifications to the material components and dosage in these embodiments also belong to the protection scope of the present invention. A detailed description of the ApNGT gene and specific application of a kind of Actinobacillus pleuropneumoniae provided by the present invention below in conjunction with accompanying drawing

[0033] Two. the bacterial classification used in the present invention and plasmid source are as follows:

[0034]

[0035] (1) After extracting the p...

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Abstract

The invention relates to specificity mutation and application of nucleotide at the 1405th bit and the 1406th bit of the ApNGT gene (NCBI:ACCESSION A3N2T3) of actinobacillus pleuropneumoniae. The nucleotide is the nucleotide shown in SEQ ID NO:1 or the nucleotide shown in SEQ ID NO:2. ApNGT is glycosylation transferase coming from actinobacillus pleuropneumoniae, can recognize the N-X-S / T sequence in polypeptide or protein, and can transfer Glucose (Glc) to an Asn residue from an activated donor UDP-Glc. After mutated ApNGT is expressed in actinobacillus pleuropneumoniae, the glycosylation efficiency at the polypeptide level is improved by 160 or more times compared with wild ApNGT, great convenience is provided for galactosylated modification of polypeptide and protein type biological products. Stability of mutated ApNGT is good, and ApNGT can be used as a tool enzyme, can be easily produced in a commercialized mode and has wide application prospects.

Description

technical field [0001] The invention belongs to the field of glycoengineering in molecular biology, and relates to specific mutations of the 1405th and 1406th nucleotides of the ApNGT gene of Actinobacillus pleuropneumoniae, and an enzyme synthesis method for rapid and simple glucosylation of polypeptides and proteins. Background technique [0002] Actinobacillus pleuropneumoniae was first reported in 1957. It was known as Haemophilus parahaemolyticus earlier. In 1964, it was officially announced as the pathogen of porcine pleuropneumonia. closely related. Belonging to Pasteurellaceae, actinomycetes, is a Gram-negative polymorphic small coccus, facultative anaerobic, with capsule, flagella and pili, can not form spores, is a respiratory parasite, mainly Exist in the lungs and tonsils of sick animals, sick pigs and pigs with bacteria are the main source of infection of this disease. [0003] Glycoprotein plays an important role in life activities, such as cell signal recogn...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N9/10C07K1/107
CPCC12N9/1051C07K1/1077
Inventor 程剑松宋其涛王鹏
Owner NANKAI UNIV
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