Difference between revisions of "Part:BBa K2886007"
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We perfect the description of VEGF-165 to make more iGEMers can know about and search for VEGF-165 expediently. And to test our STEP system, we need a large amount of VEGF-165. However, buying VEGF from companies will be really expensive. The average price of 1mg VEGF-165 is as high as 10,000 yuan (nearly 5000 dollars). Thus, we decide to construct plasmid and express VEGF-165 on ourselves. We design a sequence coding VEGF-165 followed by a 9×His tag and then entrust Sangon Biotech to synthesis it totally on the backbone pET28a. To our delighted, we succeeded in expressing VEGF-165 in E. coli and then purifying it. The result is showed in Figure 5.
 
We perfect the description of VEGF-165 to make more iGEMers can know about and search for VEGF-165 expediently. And to test our STEP system, we need a large amount of VEGF-165. However, buying VEGF from companies will be really expensive. The average price of 1mg VEGF-165 is as high as 10,000 yuan (nearly 5000 dollars). Thus, we decide to construct plasmid and express VEGF-165 on ourselves. We design a sequence coding VEGF-165 followed by a 9×His tag and then entrust Sangon Biotech to synthesis it totally on the backbone pET28a. To our delighted, we succeeded in expressing VEGF-165 in E. coli and then purifying it. The result is showed in Figure 5.
  
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[[File:Figure2 for cECFP.jpeg|600px|thumb|left|'''Figure 1:''' The gel of VEGF. The original VEGF is 38.2 kDa, and it becomes about 48 kDa after we adding the His-9 tag. On the gel, we can see a light band between 48.0 kDa and 63 kDa, which proves that we succeed in expressing and purifying VEGF-His9.]]
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[[File:T--Fudan-CHINA--figuer1for007.png|600px|thumb|left|'''Figure 1:''' The gel of VEGF. The original VEGF is 38.2 kDa, and it becomes about 48 kDa after we adding the His-9 tag. On the gel, we can see a light band between 48.0 kDa and 63 kDa, which proves that we succeed in expressing and purifying VEGF-His9.]]
 
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Revision as of 08:56, 17 October 2018


VEGF-His9


Usage and Biology

BBa_K2886007 encodes VEGF-165 with His9 tag. VEGF is a big family including lots of types. VEGF-121 was firstly introduced to iGEM by Slovenia in 2012 (BBa_K782061) and then was improved by SYSU-CHINA in 2017 (BBa_K2298001). VEGF-165 was also firstly introduced to iGEM by NYMU_Taiwan in 2012(BBa_K896922). However, there is no detail in the part wiki at all except for the part name, and actually we find this part using registry blast. This year, Fudan-CHINA makes new improvements to VEGF-165 to make it more useful and accessible to enrich the choices of iGEMers.

We perfect the description of VEGF-165 to make more iGEMers can know about and search for VEGF-165 expediently. And to test our STEP system, we need a large amount of VEGF-165. However, buying VEGF from companies will be really expensive. The average price of 1mg VEGF-165 is as high as 10,000 yuan (nearly 5000 dollars). Thus, we decide to construct plasmid and express VEGF-165 on ourselves. We design a sequence coding VEGF-165 followed by a 9×His tag and then entrust Sangon Biotech to synthesis it totally on the backbone pET28a. To our delighted, we succeeded in expressing VEGF-165 in E. coli and then purifying it. The result is showed in Figure 5.

Figure 1: The gel of VEGF. The original VEGF is 38.2 kDa, and it becomes about 48 kDa after we adding the His-9 tag. On the gel, we can see a light band between 48.0 kDa and 63 kDa, which proves that we succeed in expressing and purifying VEGF-His9.



Considering the successful results, we are willing to share this part with all iGEMers around the world to express and purify VEGF-165 on yourselves at an extremely low cost, enabling you to spare more funds for other experiments and human practice.

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.rc site found at 166