Difference between revisions of "Part:BBa K5335029"
(→‎Experimental Verification)
Line 59: Line 59:
 
<center><b>Figure 4. Confocal microscopy images of root tissues.  
 
<center><b>Figure 4. Confocal microscopy images of root tissues.  
 
<br>
 
<br>
−
(A) AmCyan fluorescence channel showing. (B) No-stain control. (C) Merged image of AmCyan fluorescence and brightfield. (D) High-magnification view of root hair cells in the merged image, showing.</b> </center>
+
(A) AmCyan fluorescence channel showing. (B) No-stain control. (C) Merged image of AmCyan fluorescence and brightfield. (D) High-magnification view of root hair cells in the merged image.</b> </center>
 
<br>
 
<br>
 
<html>
 
<html>
Line 75: Line 75:
 
</body>
 
</body>
 
</html>
 
</html>
 +
 
===Sequence and Features===
 
===Sequence and Features===
 
<partinfo>BBa_K5335029 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K5335029 SequenceAndFeatures</partinfo>
 
===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K5335029 parameters</partinfo>
 
<partinfo>BBa_K5335029 parameters</partinfo>

Revision as of 03:30, 1 October 2024


Spycatcher-CPPs(R9-Tag)

The SpyCatcher-CPPs system, designed and employed in our team project, serves as a versatile tool for validating project feasibility and aiding in the characterization of other components. Originating from the CnaB2 domain of Streptococcus pyogenes fibronectin-binding protein, this system comprises a 12 kDa SpyCatcher protein and a 13-residue SpyTag peptide. The spontaneous formation of an isopeptide bond between SpyC and SpyT enables the efficient delivery of target proteins, fused with SpyTag and the cell-penetrating peptide R9-Tag, into plant cells.

Experimental Verification

The construction and validation of the SpyCatcher component in this part involved experiments such as colony PCR, SDS-PAGE, and Western blotting. Detailed procedures for the validation can be found in the part: BBa_K5335025.


Here are some of the results: 无标题文档


Figure 1. Agarose gel electrophoresis image of colony PCR products (target band at 2200 bp)

无标题文档

Figure 2. SDS-PAGE gel electrophoresis image.

无标题文档

Figure 3. Western Blot Image.

The construction and validation of the CPPs(R9-Tag) component in this part involved the experiment,detection of plant cell penetration by AmCyan-CPPs fusion protein using laser scanning confocal microscopy. Detailed procedures for the validation can be found in the part: BBa_K5335027.


Here are some of the results: 无标题文档

Figure 4. Confocal microscopy images of root tissues.
(A) AmCyan fluorescence channel showing. (B) No-stain control. (C) Merged image of AmCyan fluorescence and brightfield. (D) High-magnification view of root hair cells in the merged image.

无标题文档

Figure 5. Confocal fluorescence microscopy images of protoplasts prepared from Arabidopsis roots.
(A) AmCyan fluorescence channel showing. (B) No-stain control. (C) Merged image of AmCyan fluorescence and brightfield.

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
    COMPATIBLE WITH RFC[1000]

Functional Parameters