Abstract
【Objective】 The Pcpme3 gene of Phytophthora capsici was expressed in Eukaryotic expression system to obtain purified protein and specific antiserum, and to detect the function of Pcpme3 in phytophthora-infected pepper through Western blot. 【Method】 Mature peptide of Pcpme3 obtained by RT-PCR was cloned into pPIC9K, and the eucaryotic expression plasmid was constructed. The recombinant plasmid was transformed into Pichia pastoris GS115 by electroporation. After induced by methanol, the culture supernatant was analyzed by SDS-PAGE, and the antiserum was prepared. 【Result】 The high copy transformants with Mut+/His+ phenotype were selected by screening on MD, MM and G418 medium and PCR. The culture supernatant was analyzed by SDS-PAGE after induced by 1% methanol, and the results showed that a specific protein of about 43 kD was expressed. The specific antiserum was produced after the rabbit was immunized with recombinant protein, and Western blot indicated that Pcpme3 could be highly expressed in phytophthora-infected peppers, and the expressed level became stronger with the symptom became heavier. 【Conclusion】 Antiserum prepared from expressed protein can be used to detect the function of Pcpme3 in the pathogenesis of Phytophthora capsici.
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Claims 6
Claim · model-written summary
High copy transformants with Mut+/His+ phenotype were selected.
- Taxon
- Pichia pastoris
.jpg?width=60)
Komagataella pastorise1 - Outcome
- high copy transformants with Mut+/His+ phenotypee0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
.jpg?width=500)
Verbatim
e0“The high copy transformants with Mut+/His+ phenotype were selected by screening on MD, MM and G418 medium and PCR.”
e1“The recombinant plasmid was transformed into Pichia pastoris GS115 by electroporation.”
Claim · model-written summary
Induction with 1% methanol yielded a specific protein of about 43 kD.
- Taxon
- Pichia pastorisKomagataella pastorise1
- Treatment
- 1% methanole0
- Outcome
- a specific proteine0
- Value
- about 43 kDe0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The culture supernatant was analyzed by SDS-PAGE after induced by 1% methanol, and the results showed that a specific protein of about 43 kD was expressed.”
e1“The recombinant plasmid was transformed into Pichia pastoris GS115 by electroporation.”
Claim · model-written summary
Specific antiserum was produced after rabbit immunization with recombinant protein.
- Taxon
- rabbit
.jpg?width=60)
Oryctolagus cuniculuse0 - Outcome
- specific antiserume0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
.jpg?width=500)
Verbatim
e0“The specific antiserum was produced after the rabbit was immunized with recombinant protein”
Claim · model-written summary
Pcpme3 was highly expressed in infected peppers.
- Interactor
- phytophthora
Phytophthorae0pathogen - Gene
- Pcpme3e0
- Outcome
- expressede0
- Direction
- highlye0
CapsicumOrganisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Western blot indicated that Pcpme3 could be highly expressed in phytophthora-infected peppers”
Claim · model-written summary
Pcpme3 expression became stronger as pepper symptoms worsened.
- Interactor
- phytophthoraPhytophthorae0pathogen
- Gene
- Pcpme3e0
- Outcome
- expressed levele0
- Direction
- strongere0
- Qualifier
- the symptom became heaviere0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Western blot indicated that Pcpme3 could be highly expressed in phytophthora-infected peppers, and the expressed level became stronger with the symptom became heavier.”
Claim · model-written summary
The authors conclude that the antiserum can detect Pcpme3 function in Phytophthora capsici pathogenesis.
- Taxon
- Phytophthora capsici
.jpg?width=60)
Phytophthora capsicie0 - Gene
- Pcpme3e0
- Outcome
- detect the function of Pcpme3e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
.jpg?width=500)
Verbatim
e0“Antiserum prepared from expressed protein can be used to detect the function of Pcpme3 in the pathogenesis of Phytophthora capsici.”