Recombinant vector used for phellinus igniarius (L.ex Fr.) Quel. [Fomes igniarius (L.) Fr. fruiting body genetic expression, construction method for recombinant vector, and recombinant vector genetic transformation method

A genetic transformation method and genetic expression technology, applied in the direction of microorganism-based methods, the use of vectors to introduce foreign genetic material, recombinant DNA technology, etc., can solve problems such as the inability to establish a Phellinus genetic transformation system, achieve fast speed, and have good application prospects , the effect of simplifying the operation steps

Active Publication Date: 2020-11-10
NORTHEAST FORESTRY UNIVERSITY
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  • Claims
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Problems solved by technology

[0006] Based on the above deficiencies, the purpose of the present invention is to provide a kind of recombination binary vector and construction method for the genetic expression of Phellinus Phellinus. Solved the problem that the genetic transformation system of Phellinus chinensis could not be established

Method used

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  • Recombinant vector used for phellinus igniarius (L.ex Fr.) Quel. [Fomes igniarius (L.) Fr. fruiting body genetic expression, construction method for recombinant vector, and recombinant vector genetic transformation method
  • Recombinant vector used for phellinus igniarius (L.ex Fr.) Quel. [Fomes igniarius (L.) Fr. fruiting body genetic expression, construction method for recombinant vector, and recombinant vector genetic transformation method
  • Recombinant vector used for phellinus igniarius (L.ex Fr.) Quel. [Fomes igniarius (L.) Fr. fruiting body genetic expression, construction method for recombinant vector, and recombinant vector genetic transformation method

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[0057] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are all purchased from conventional biochemical reagent stores unless otherwise specified.

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Abstract

The invention discloses a recombinant vector used for phellinus igniarius (L.ex Fr.) Quel. [Fomes igniarius (L.) Fr. fruiting body genetic expression, a construction method for the recombinant vector,and a recombinant vector genetic transformation method. The sequence of the vector is disclosed by SEQ ID No:13. The construction method comprises the following step of replacing a CaMV35S promoter of a binary vector pCAMBIA-1301 with a phellinus igniarius (L.ex Fr.) Quel. [Fomes igniarius (L.) Fr. fruiting body homologous promoter to obtain a binary vector suitable for the phellinus igniarius (L.ex Fr.) Quel. [Fomes igniarius (L.) Fr. fruiting body. The recombinant vector genetic transformation method comprises the following steps of: utilizing a freeze thawing method to convert the recombinant binary vector into agrobacterium tumefaciens, and obtaining positive bacteria; utilizing the agrobacterium tumefaciens to infect phellinus igniarius (L.ex Fr.) Quel. [Fomes igniarius (L.) Fr. hyphae, carrying out co-culture on a culture medium, and subsequently, covering a low melting point PDA culture medium; and then, transferring bacterial colonies to a new culture medium, extracting DNA from the bacterial colony which grows luxuriantly, and utilizing a PCR technology to identify a positive bacterial strain. The recombinant vector has the advantages of being simple in operation, high ina conversion rate and the like, the phellinus igniarius (L.ex Fr.) Quel. [Fomes igniarius (L.) Fr. fruiting body genetic expression is successfully realized, homologous knockout, overexpression and heterologous expression in the phellinus igniarius (L.ex Fr.) Quel. [Fomes igniarius (L.) Fr. fruiting body become possible, and the problem that a phellinus igniarius (L.ex Fr.) Quel. [Fomes igniarius(L.) Fr. fruiting body genetic transformation system can not be established is solved.

Description

technical field [0001] The invention belongs to the field of genetic breeding, and in particular relates to a recombinant vector for genetic expression of Phellinus linteus, a construction method and a genetic transformation method. Background technique [0002] Phellinus linteus has a history of more than 2,000 years as a famous traditional medicinal fungus in China. According to "Compendium of Materia Medica", Phellinus linteus can be used for antidiarrheal, metrorrhagia, diarrhea due to spleen deficiency, in addition to detoxification, blood circulation, and five internal organs. Modern research has shown that Phellinus linteus has the functions of regulating immunity, inhibiting the proliferation of cancer cells, and inducing apoptosis of cancer cells. It is currently one of the internationally recognized medicinal fungi with better anti-cancer effects. Triterpenes, flavonoids and phenolics. [0003] At present, the research on the medicinal components of Phellinus lin...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/80C12N15/66C12N15/65C12N1/15C12Q1/686C12Q1/04C12R1/645
CPCC12N15/80C12N15/66C12N15/65C12Q1/686C12Q2531/113Y02A50/30
Inventor 邹莉王旭彤孙婷婷许昕薷孙健曲晓磊王世新刘增才尹龙川
Owner NORTHEAST FORESTRY UNIVERSITY
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