Difference between revisions of "Part:BBa K4653105"

 
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Protein phosphomutase BvEP from Bacillus velezensis LJ02, has been shown being able to induce the PTI and ETI immune pathways in tomato fruits, stimulate ROS outbreak and related enzyme expression, and have no significant effect on the weight and nutrient content of tomato fruits.
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BvEP is an elicitor protein phosphopentomutase from <I>Bacillus velezensis</I> LJ02 which improves the tomato resistance to <I>B. cinerea</I>. High purity BvEP proteins is shown to induce the hypersensitivity response (HR) in <I>Nicotiana tabacum</I>. BvEP reduced the rotting rate and lesion diameter of tomato fruits caused by B. cinerea, and increased the expression of Pattern-triggered Immunity (PTI), Effector-triggered Immunity (ETI), systemic acquired resistance (SAR) related genes, ROS content, SOD and POD activities in tomato fruits, while there was no significant effect on weight loss and VC contents of tomato fruits.  
  
 
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===Biology===
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BvEP is an elicitor protein phosphopentomutase from <I>Bacillus velezensis</I> LJ02. Phosphopentomutase (PPM) is found in bacterial and mammalian cells and is responsible for the interconversion of ribose-5-phosphate (R5P) and ribose-1-phosphate (R1P). <I>B. velezensis</I>  LJ02, as a beneficial microorganism. The srcretor BvEP can activate PTI and ETI reactions of tomato and improve the activity of peroxides and superoxide dismutase, and finally enhance the resistance of tomato to <I>B. cinerea</I>.
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===Design===
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The fragment was amplified from the total DNA of <I>B. Velezensis</I> LJ02 by specific primers. We retrieved the total genome of Bacillus Velez and entered the primer sequence after opening its FASTA file on Snapgene.
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<center><html><img src="https://static.igem.wiki/teams/4653/wiki/parts/szu-parts-k4653105-1.jpg" width="1280" height="192"  /></html></center>
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<center><b>Figure 1. The location of the primers provided in the literature amplified on the total genome</b></center>
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The fragments simulated by primers were used as BvEP sequences, which were blasted and showed high homology among the bacillus.  However, we found that the fragment amplified by the primer was incomplete and lacked the start codon, so we performed blastp on the incomplete sequence in NCBI to obtain the complete amino acid sequence. Finally, the complete sequence of the protein was obtained on the whole genome.
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===Plasmid construction===

Revision as of 16:14, 8 October 2023


BvEP: Protein phosphomutase

BvEP is an elicitor protein phosphopentomutase from Bacillus velezensis LJ02 which improves the tomato resistance to B. cinerea. High purity BvEP proteins is shown to induce the hypersensitivity response (HR) in Nicotiana tabacum. BvEP reduced the rotting rate and lesion diameter of tomato fruits caused by B. cinerea, and increased the expression of Pattern-triggered Immunity (PTI), Effector-triggered Immunity (ETI), systemic acquired resistance (SAR) related genes, ROS content, SOD and POD activities in tomato fruits, while there was no significant effect on weight loss and VC contents of tomato fruits.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 115


Biology

BvEP is an elicitor protein phosphopentomutase from Bacillus velezensis LJ02. Phosphopentomutase (PPM) is found in bacterial and mammalian cells and is responsible for the interconversion of ribose-5-phosphate (R5P) and ribose-1-phosphate (R1P). B. velezensis LJ02, as a beneficial microorganism. The srcretor BvEP can activate PTI and ETI reactions of tomato and improve the activity of peroxides and superoxide dismutase, and finally enhance the resistance of tomato to B. cinerea.

Design

The fragment was amplified from the total DNA of B. Velezensis LJ02 by specific primers. We retrieved the total genome of Bacillus Velez and entered the primer sequence after opening its FASTA file on Snapgene.

Figure 1. The location of the primers provided in the literature amplified on the total genome


The fragments simulated by primers were used as BvEP sequences, which were blasted and showed high homology among the bacillus. However, we found that the fragment amplified by the primer was incomplete and lacked the start codon, so we performed blastp on the incomplete sequence in NCBI to obtain the complete amino acid sequence. Finally, the complete sequence of the protein was obtained on the whole genome.

Plasmid construction