Difference between revisions of "Part:BBa K1859015"
| Line 21: | Line 21: | ||
<br> | <br> | ||
| − | <p>We improved the | + | <p>We improved the 3´side of the intron |
| − | <a href= "https://parts.igem.org/wiki/index.php?title=Part: | + | <a href= "https://parts.igem.org/wiki/index.php?title=Part:BBa_K1332005" > [BBa_K1332005] |
</a> of iGEM2014 and made [BBa_K1859015]. | </a> of iGEM2014 and made [BBa_K1859015]. | ||
Circular parts of iGEM2014 are cloned only sequence of ribozyme involved the splicing from td intron in T4 phage. | Circular parts of iGEM2014 are cloned only sequence of ribozyme involved the splicing from td intron in T4 phage. | ||
However, [BBa_K1859015] is cloned the sequence except the ribozyme extra in addition to | However, [BBa_K1859015] is cloned the sequence except the ribozyme extra in addition to | ||
| − | <a href= "https://parts.igem.org/wiki/index.php?title=Part: | + | <a href= "https://parts.igem.org/wiki/index.php?title=Part:BBa_K1332005" > [BBa_K1332005] |
</a> | </a> | ||
. | . | ||
| Line 32: | Line 32: | ||
<p> | <p> | ||
| − | We attached this part[BBa_1859015] to | + | We attached this part[BBa_1859015] to 5'sides of |
His-RFP without stop codon | His-RFP without stop codon | ||
<a href= "https://parts.igem.org/wiki/index.php?title=Part:BBa_K1332002" > [BBa_K1332002] | <a href= "https://parts.igem.org/wiki/index.php?title=Part:BBa_K1332002" > [BBa_K1332002] | ||
</a>. | </a>. | ||
| − | Then, it was connected to | + | Then, it was connected to 5'sides of |
| − | <a href= "https://parts.igem.org/wiki/index.php?title=Part: | + | <a href= "https://parts.igem.org/wiki/index.php?title=Part:BBa_K1859016" > [BBa_K1859016] |
</a>. | </a>. | ||
In addition,Lacl | In addition,Lacl | ||
Revision as of 08:48, 22 September 2015
the 3´side of the intron[BBa_K1332005] +complementary seqence
[BBa_K1859015] can let mRNA circularize efficiently in E. coli by combining it with [BBa_K1859016] .
Circular Parts, the 3'side of the intron [BBa_K1332005] and the 5'side of the intron [BBa_K1332003] , is what use sequence subcloned ribozyme involved self‐splicing in T4 Phage. By associating Circular Parts on both ends of optional sequence, you can circularize mRNA making its DNA.
As for details of circular mRNA, refer to ‘iGEM Gifu 2014' .
We improved the 3´side of the intron [BBa_K1332005] of iGEM2014 and made [BBa_K1859015]. Circular parts of iGEM2014 are cloned only sequence of ribozyme involved the splicing from td intron in T4 phage. However, [BBa_K1859015] is cloned the sequence except the ribozyme extra in addition to [BBa_K1332005] .
We attached this part[BBa_1859015] to 5'sides of His-RFP without stop codon [BBa_K1332002] . Then, it was connected to 5'sides of [BBa_K1859016] . In addition,Lacl [BBa_R0010] , RBS [BBa_B0034] and DT [BBa_B0015] was conbined with it, making [BBa_K1859026] . [BBa_K1859026] is the generator to improve circular efficiency of mRNA. The sequence that cloned extra in [BBa_K1859015] has complementarity with the sequence that cloned extra in [BBa_K1859016].
You can let mRNA circularize efficiently by using [BBa_K1859026] ([BBa_K1859015] and [BBa_K1859016] ) than using the generator of 2014 [BBa_K1332011] ( [BBa_K1332005] and [BBa_K1332003] ).