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Method of cloning Phytophthora parasitica effector

Technology of an effector, blackleg bacteria, applied in the field of biology

Active Publication Date: 2019-07-05
TOBACCO RES INST CHIN AGRI SCI ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of these effectors come from Phytophthora infestans and Phytophthora sojae, while the research on RxLR effectors of tobacco blackleg has not been reported yet

Method used

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  • Method of cloning Phytophthora parasitica effector
  • Method of cloning Phytophthora parasitica effector
  • Method of cloning Phytophthora parasitica effector

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Experimental program
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Embodiment Construction

[0018] The present invention will be described in detail below in combination with specific embodiments.

[0019] Cloning of Effect Factor HJ01 of Blackleg Fungus

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Abstract

The invention discloses a method of cloning Phytophthora parasitica effector. The method comprises: designing primers; extracting Phytophthora parasitica genome DNA; performing PCR (polymerase chain reaction) amplification on tobacco Phytophthora parasitica effector gene; recycling and purifying the PCR product; preparing Escherichia coli DH5lapha competence; linking the amplification product withpMDTM 19-T Vector, and converting Escherichia coli DH5alpha; converting Escherichia coli; screening a positive clone, and performing PCR identification on a bacterial liquid; subjecting the screenedpositive clone to sequencing identification to obtain the Phytophthora parasitica effector. The method has the advantages that the Phytophthora parasitica effector is cloned and its functionality is analyzed.

Description

technical field [0001] The invention belongs to the technical field of biology, and relates to the cloning and functional analysis of the blackleg pathogen effect factor HJ01. Background technique [0002] Tobacco black shank belongs to Stramenopila or Chromista, Omycota, Oomycetes, Peronosporales, Pythiaceae, Phytophthora Genus (Phytophthora). Plant pathogenic bacteria can secrete effector proteins into the host, allowing them to suppress the plant's immune system, disrupt cell activity, and eventually cause disease (Kazan & Lyons, 2014). Bioinformatics analysis showed that pathogenic oomycetes encode a large class of effector proteins, namely RxLR effector proteins (Arginine-any amino acid-Leucine-Argininemotif, RxLR). The N-terminus of RxLR effector protein contains a conserved Arginine-any amino acid-Leucine-Arginine motif domain (R, arginine; x, any amino acid; L, leucine). The RxLR motif is usually followed by a small conserved Glutamic acid-Glutamic acid-Arginine (...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12N15/31C12N15/70
CPCC07K14/37C12N15/10C12N15/70Y02A50/30
Inventor 王文静王凤龙陈德鑫任广伟焦芳婵王静
Owner TOBACCO RES INST CHIN AGRI SCI ACAD
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