Composite

Part:BBa_K4586033

Designed by: Mohamed Mohamed Saad Aboelghare   Group: iGEM23_AFCM-Egypt   (2023-10-12)
Revision as of 14:08, 12 October 2023 by E m1 (Talk | contribs) (→‎Experimental Characterization)

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Anti CD19-Exosomes receptor

Usage and Description

This composite parts codes for lamp2b which is a transmembrane protein known as lysosome-associated membrane glycoprotein 2b, which is unique to the membrane of exosomes and its external domain conjugated to ANTI CD19 to mediate its expression on the surface of the exosomes.

Experimental Characterization

In order to amplify this DNA part, we used PCR amplification to reach the desired concentration to complete our experiments using specific forward and reverse primers, running the parts on gel electrophoresis as this part presents lane (p5) including lamb2p an anti-CD19 , and then we measured the specific concentration of the running part using Real-Time PCR as shown in the following figure.





We performed the double digestion method for this part in the prefix and suffix with its specific restriction enzyme and applied this part to gel electrophoresis as shown in the following figure lane(P5).



After the ligation step, we did a culture of the ligated product to specifically select the optimum colonies to screen it using Colony PCR to make sure that our parts were correctly ligated in the plasmid. This figure shows the Cell culture plate of transformed pCDNA vector 2 containing insert parts. This plasmid contains Loading system(CD63-L7Ae)-exosomal receptor(LAMP2B-anti-CD19)-connexin(CX43)




Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 614
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1070


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