Difference between revisions of "Part:BBa K2886007"

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

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2886007 short</partinfo>
-We present VEGF-His9 as our best basic part to enable iGEMers to express and purify VEGFA-165
-easily.
-VEGF (vascular endothelial growth factor), originally known as VPF (vascular permeability factor), is a ligand commonly used in research. As a kind of endogenous growth factor produced by cells humans and mice, VEGF has various biological effects such as stimulates the formation of blood vessels and enhance the permeability of blood vessels. VEGF is a big family, and there are a lot of types of VEGF including VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E and PGF, of which VEGF-121 and VEGF-165 are two main types in body and research.
-VEGF-121 was introduced to iGEM by Slovenia in 2012 (BBa_K782061) and then was improved by SYSU-CHINA in 2017 (BBa_K2298l002). 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, and we actually 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.
+===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 perfected the description of VEGF-165 to make more iGEMers can know about and search for VEGF-165 expediently. 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 designed a sequence coding VEGF-165 followed by a 9×His tag and then entrusted Sangon Biotech to synthesis it totally on the backbone [[User:Vesalius|Vesalius]] ([[User talk:Vesalius|talk]]) 19:10, 14 October 2018 (UTC). To our delighted, we succeeded in expressing VEGF-165 in E. coli and then purifying it. Here are the result.;
+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.
-From the gel, we can see that19:10, 14 October 2018 (UTC)19:10, 14 October 2018 (UTC)19:10, 14 October 2018 (UTC)19:10, 14 October 2018 (UTC)
+[[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.]]
+<br style="clear: both" />
-Considering the successful results, we are willing to share this part with all iGEMer 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.
+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.
-<!-- Add more about the biology of this part here
-===Usage and Biology===
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