Team:AHUT China/Project/Biobrick
Biobrick
Our submission parts
<2-6-8> is the shortest pathway obtained from the experiment in our project. It is made up of 5 DNA single strands, which are Site NO.2, Site NO.6, Site NO.8 and Line <2,6>, Line <6,8> obtained from bridging-PCR. We designed two restriction enzyme digestion cutting site respectively in the primers of Site NO.2 and Site NO.8 in order to insert plasmid pSB1C3.Every site contains 40bp DNA sequence at random and every adjacent site is connected by line, which adds up to 13 lines. Lines are represented by 52bp DNA single strand, with post-20 complementary base pairs from the first and pre-20 complementary base pairs from the last site plus 12bp information bits in the middle. The NO.21 base to NO.32 base in the 40bp of pathway <2,6> and <2,8> represents the information sequence. The pre-4bp sequences contain the information of identification point, the mid-4bp sequences contain the information of the specified line, and the post-4bp contain the information of the path length. Through DNA Gel electrophoresis and Sequencing analysis, <2-6-8> proved to be the shortest pathway. The calculated actual length of <2-6-8> is 26 mile in the reality.
<2-4-8> is another part obtained in the experiment of our project, which has the same base length as <2-6-8>, the shortest pathway.We designed two restriction enzyme digestion cutting site respectively in the primers of Site NO.2 and Site NO.8 in order to insert plasmid pSB1C3.Every site contains 40bp DNA sequence at random and every adjacent site is connected by line, which adds up to 13 lines. Lines are represented by 52bp DNA single strand, with post-20 complementary base pairs from the first and pre-20 complementary base pairs from the last site plus 12bp information bits in the middle. The NO.21 base to NO.32 base in the 40bp of pathway <2,4> and <4,8> represents the information sequence. The pre-4bp sequences contain the information of identification point, the mid-4bp sequences contain the information of the specified line, and the post-4bp contain the information of the path length. Through DNA Gel electrophoresis and Sequencing analysis, <2-6-8> proved to be the shortest pathway. The calculated actual length of <2-6-8> is 28 mile in the reality.
Improved part
Solution24578 is the shortest pathway in the project of 2014 of AHUT-China, which Node NO.5 is a necessary node to be passed in order to get to Node NO.8. However, in our project of this year, we don’t have to pass Site NO.5 while taking Site NO.2 as the start point and Site NO.8 as the terminal, which provides us with more diverse pathway choices and more conforms to the real situation. In our project of 2016, the DNA single strands of the sites and lines are connected by bridging-PCR. 12 information bits are added to the DNA single strand of the lines, which the pre-4bp sequences contain the information of identification point, the mid-4bp sequences contain the information of the specified line, and the post-4bp contain the information of the path length. In order to obtain Solution24578, it needs to construct recombinant plasmid in every possible pathway. And then the recombinant plasmid was inserted in E.coli, where the fluorescent protein gene in Node NO.5 get expressed. Thus, all the pathways passing Node NO.5 screened out. By contrast, we don’t have to go through a series of complex process like what have been depicted above in order to obtain the shortest pathway <2-6-8> in this year.
More information please see our submission parts form:
| Name | Type | Description | Length |
|---|---|---|---|
| BBa_K2044000 | DNA | Solution <2-6-8> | 144bp |
| BBa_K2044001 | DNA | Solution <2-4-8> | 144bp |
| BBa_K2044002 | DNA | Line<2,1> | 52bp |
| BBa_K2044003 | DNA | Line<2,3> | 52bp |
| BBa_K2044004 | DNA | Line<2,4> | 52bp |
| BBa_K2044005 | DNA | Line<2,6> | 52bp |
| BBa_K2044006 | DNA | Line<3,6> | 52bp |
| BBa_K2044007 | DNA | Line<4,5> | 52bp |
| BBa_K2044008 | DNA | Line<4,6> | 52bp |
| BBa_K2044009 | DNA | Line<4,8> | 52bp |
| BBa_K2044010 | DNA | Line<5,7> | 52bp |
| BBa_K2044011 | DNA | Line<6,4> | 52bp |
| BBa_K2044012 | DNA | Line<6,8> | 52bp |
| BBa_K2044013 | DNA | Line<7,8> | 52bp |
| BBa_K2044014 | DNA | Line<1,4> | 52bp |
| BBa_K2044015 | DNA | Site No.1 | 40bp |
| BBa_K2044016 | DNA | Site No.2 | 40bp |
| BBa_K2044017 | DNA | Site No.3 | 40bp |
| BBa_K2044018 | DNA | Site No.4 | 40bp |
| BBa_K2044019 | DNA | Site No.5 | 40bp |
| BBa_K2044020 | DNA | Site No.6 | 40bp |
| BBa_K2044021 | DNA | Site No.7 | 40bp |
| BBa_K2044022 | DNA | Site No.8 | 40bp |
