Difference between revisions of "Part:BBa K2886007"

Latest revision as of 20:33, 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(Part: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.

Improvement on a previous part

VEGF-165 was firstly introduced to iGEM by NYMU_Taiwan in 2012(Part: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.

For more detailed information, please click herehttp://2018.igem.org/Team:Fudan-CHINA/Improve to read our wiki.

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

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 <partinfo>BBa_K2886007 short</partinfo>
-VEGF (vascular endothelial growth factor) is a common ligand used in synthetic biology. And there are a lot of types of VEGF in human and rice like VEGFA-121, VEGFA-165 and so on. And in human, there are two main types of VEGFA: VEGF-121 and VEGFA-165.
-VEGFA-121 was firstly introduced to iGEM by  iGEM12_Slovenia in 2012, which was from mouse. In 2017, iGEM17_SYSU-CHINA improved this part by isolating it from the initial composite part and submitting the counterpart in human, making VEGFA-121 more mobile and suitable for human cell lines.
-VEGFA-165 also was firstly introduced to iGEM by  iGEM12_NYMU_Taiwan in 2012(BBa_K896922). However, there is no detail in the part wiki at all, and we actually find this part through blast tools. This year, Fudan-CHINA makes new improvements to VEGFA to make it more useful  and enrich the chioces of iGEMers.
+===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([[Part: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.
-According to literatures, VEGFA-165 has a higher affinity to heparin and might has higher activity than VEGFA-121. Besides, VEGFA-165 is much more adopted in research compared with VEGFA-121. And what's more, buying VEGFA-165 from companies might be very expensive, so we add a His tag that could help us to express and purify VEGFA-165 easily. And We did succeed in collecting purified VEGFA-165 with the part we constructed, proving its reliability. What is also worth mentioning is that we could also us GS tag, but given it's even longer than VEGF itself, we turned to His tag ultimately.
+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.
-All in all, through our work, VEGFA becomes much more available and easy to use.
+[[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.]]
+<br style="clear: both" />
-<!-- Add more about the biology of this part here
-===Usage and Biology===
+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.
+===Improvement on a previous part===
+VEGF-165 was firstly introduced to iGEM by NYMU_Taiwan in 2012([[Part: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.
+For more detailed information, please click here[[http://2018.igem.org/Team:Fudan-CHINA/Improve]] to read our wiki.
 <!-- -->