Thermophilic streptomyces hygroscopicus cDNA library construction, and expression and expression methods of two greenhouse farmer's lungpathopoiesia antigen polypeptides

A technology of Streptomyces hygroscopicus and antigenic polypeptides, applied in recombinant DNA technology, chemical instruments and methods, botanical equipment and methods, etc., can solve problems such as aggravating the economic burden of sick farmers and social burden, increasing diagnosis and treatment costs, and delaying treatment timing.

Inactive Publication Date: 2014-01-01
中国医科大学附属第四医院
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Problems solved by technology

Due to the lack of awareness of greenhouse lung disease by doctors in grassroots hospitals, they are often misdiagnosed as common colds and asthma, which delays the best time for treatment, resulti

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  • Thermophilic streptomyces hygroscopicus cDNA library construction, and expression and expression methods of two greenhouse farmer's lungpathopoiesia antigen polypeptides
  • Thermophilic streptomyces hygroscopicus cDNA library construction, and expression and expression methods of two greenhouse farmer's lungpathopoiesia antigen polypeptides
  • Thermophilic streptomyces hygroscopicus cDNA library construction, and expression and expression methods of two greenhouse farmer's lungpathopoiesia antigen polypeptides

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[0038] The constructed cDNA library of Streptomyces thermophilus hygroscopicus obtained 978 sense sequences and 347 single genes, among which the homology of one of the two genes to the outer membrane protein of Actinomyces pleuropneumoniae was 51%. The length of the open reading frame (ORF) was 1554 bp, encoding 517 amino acids; the homology of another gene with transferrin B was 42%, the length of the open reading frame (ORF) was 726 bp, encoding 241 amino acids.

[0039] The expression of the two greenhouse lung pathogenic antigen polypeptides is to subclone the two segments of the gene into a prokaryotic expression vector, and the relative molecular masses of the IPTG-induced expression products are 63,000 and 30,000 polypeptides, respectively.

[0040] The method for constructing cDNA library of Streptomyces thermophilus hygroscopicus and expressing two greenhouse lung pathogenic antigen polypeptides comprises the following steps:

[0041] 1. Construction of cDNA library ...

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Abstract

The invention provides thermophilic streptomyces hygroscopicus cDNA library construction, and expression and expression methods of two greenhouse farmer's lung pathopoiesia antigen polypeptides. The problem needed solving is that: the thermophilic streptomyces hygroscopicus cDNA library is constructed, two greenhouse farmer's lung pathopoiesia virulence genes with virulence effects need screening out for cloning in vitro and inducing expression of the two greenhouse farmer's lung pathopoiesia antigen polypeptides. The key point of construction of the thermophilic streptomyces hygroscopicus cDNA library comprises: acquiring 978 sense sequences and 347 single genes, wherein the homology of two gene fragments respectively with actinobacillus pleuropneumoniae outer membrane protein and transferrin B is 51% and 42%, and the open reading frame length of the two gene fragments respectively is 1554 bp and 726 bp, and 517 and 241 amino acids are respectively encoded. The applications of the thermophilic streptomyces hygroscopicus cDNA library comprise that: from the thermophilic streptomyces hygroscopicus cDNA library, the two greenhouse farmer's lung pathopoiesia virulence genes with virulence effects are screened out for cloning in vitro and inducing expression of the two greenhouse farmer's lung pathopoiesia antigen polypeptides, and theoretical basis and practical basis are provided for subsequent development of new diagnosis methods for farmer's lung diseases.

Description

technical field [0001] The invention belongs to the field of biological detection, and specifically relates to the construction of a Streptomyces thermophilic hygroscopicus cDNA library, the sequencing of expressed sequence tags (EST), the acquisition of two pathogenic genes that cause allergic reactions in the body through biological information means, and the screening of virulent genes. The two pathogenic genes were cloned in vitro and induced the expression of two greenhouse lung pathogenic antigen polypeptides, which provided a theoretical and practical basis for the subsequent development of new diagnostic methods for farmer's lung. Background technique [0002] 1. Current status of incidence and detection of lung diseases in farmers [0003] Farmer's lung disease is an exogenous allergic alveolitis caused by inhalation of antigenic organic dust (such as moldy hay or other moldy plant dust, etc.), usually after exposure to various moldy plants It occurs among farmers,...

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Application Information

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IPC IPC(8): C40B40/08C40B50/06C12N15/31C12N15/70C07K14/36
Inventor 王笑歌王玲玲刘朔
Owner 中国医科大学附属第四医院
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