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Method for authenticating plasmodiophora brassicae woron secretory protein

A technology of Plasmodium brassica secreting proteins, applied in the biological field, can solve problems such as limiting the research on pathogenic molecular mechanisms, inability to grow on artificially synthesized media, and lack of research reports on the identification of secreted proteins

Pending Publication Date: 2019-08-09
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Plasmodium brassicae is an obligate biotrophic organism, and currently it cannot grow on synthetic media. The unique lifestyle largely limits the research on its pathogenic molecular mechanism; more and more studies It shows that the secreted protein of the pathogen plays an important role in its pathogenic process, but there is still a lack of research reports on the identification of the secreted protein of Plasmodium brassicae

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  • Method for authenticating plasmodiophora brassicae woron secretory protein
  • Method for authenticating plasmodiophora brassicae woron secretory protein
  • Method for authenticating plasmodiophora brassicae woron secretory protein

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Embodiment 1

[0032] A method for identifying proteins secreted by Plasmodium brassicae, the specific steps are as follows:

[0033] (1)芸薹根肿菌分泌蛋白的鉴定:将氨基酸序列(MQSWLALCTTVLLAWSCSGACNCTLDVAQVCGSDGVTYRNSCLALCSGAAVAKAGPCSCTCQPSSPSDTVCGTDGRAHKSACEAECNSVKVAYTGSCIQPCSSSDAATTSMPVCGNDGQTYPNPCEARNKGILISSQGACSGQPVKCDCPDQVAPVCASDGSTYSSSCKAKCTGVSIVHLGACTN;SEQ ID NO.1)输入框内,利用SingalP 4.1在线预测(http: / / www.cbs.dtu.dk / services / SignalP / ) Signal peptide (signal peptide, SP) sequence (the amino acid sequence of the predicted signal peptide is MQSWLALCTTVLLAWSCSG; SEQ ID NO.2; the nucleic acid sequence corresponding to the signal peptide is ATGCAGAGCTGGCTTGCGCTGTGTACGACGGTTCTGCTGGCATGGTCGTGCTCCGGC; SEQ ID NO.3), select the default parameters such as figure 1 As shown, the signal peptide of PbCe1 is predicted with SignalP 4.1 default parameters ( figure 2 ),From figure 2 It can be seen from that, SignalP4.1 predicts that the signal peptide of PbCe1 is located at amino acid residues 1-19 of the amino terminal. SMA...

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Abstract

The invention discloses a method for authenticating plasmodiophora brassicae woron secretory protein. The method includes the steps of predicting a signal peptide sequence of the plasmodiophora brassicae woron secretory protein, establishing a plasmodiophora brassicae woron secretory protein carrier, extracting root RNA of arabidopsis thaliana infected with clubroot, reversely transcribing the root RNA into cDNA to serve as a PCR amplification template, amplifying the predicted signal peptide sequence through high-fidelity enzyme KOD-FX NEO, establishing pSUC2-PbCe1, conducting yeast transformation, and conducting PCR verification of a yeast transformant. By means of the method for authenticating the plasmodiophora brassicae woron secretory protein, the powerful technological support can be provided for the authentication and related subsequent research of the plasmodiophora brassicae woron secretory protein.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for identifying secretory proteins of Plasmodium brassicae. Background technique [0002] Clubroot disease is a worldwide soil-borne disease caused by Plasmodiophora brassicae Woronin, which has a wide range of hosts and can harm cabbage, cauliflower, Chinese cabbage, pak choy, rapeseed, shepherd's purse, etc. A variety of cruciferous cultivated and wild plants. Clubroot damages an area of ​​about 3.2-4 million hm per year 2 , the area of ​​occurrence and damage in a pandemic year can reach 9 million hm 2 , the average yield loss reaches 20%-30%, and the direct yield loss of the seriously ill field reaches more than 60%. In addition to directly reducing crop yields, the dormant spores released from the diseased roots are extremely vigorous and can survive in the soil for up to 20 years. Once the field is polluted by clubroot, the field will be infected for a long time and...

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Application Information

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IPC IPC(8): C12N15/81C12N15/31
CPCC12N15/81C07K14/37Y02A50/30
Inventor 余方伟王神云李建斌张伟唐君王红于利
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES