Difference between revisions of "Part:BBa K883503"
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<partinfo>BBa_K883503 short</partinfo> | <partinfo>BBa_K883503 short</partinfo> | ||
| − | This part contains TuYV coat protein and part of readthrough with His-Tag on the N terminus. Because There are iGEN illegal sites in TuYV readthrough part, we just isolate part of the readthrough, which | + | This part contains TuYV coat protein and part of readthrough with His-Tag on the N terminus. Because There are iGEN illegal sites in TuYV readthrough part, we just isolate part of the readthrough, which does not contain the illegal sites. |
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| + | Turnip Yellows Virus (TuYV) is member of the genus Polerovirus and family Luteoviridae, which is positive sense RNA virus. The viral gene encoding the coat protein and part of the readthrough for TuYV, (GenBank: NC_003743.1 (3483..5495)) was obtained from a plasmid encoding the entire viral genome (GenBank: X13063.1). With designed primers, iGEM prefix and His-tag reporter protein was added to the 5 prime end and suffix were added 3 prime end of the this construct gene separately. | ||
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| + | Then the PCR products were digested, ligated to pSB1C3 backbone, and transformed into DH5 alpha competent E.coli cells. Because we used white and red screening, the white colonies, which are colonies with right inserts, were checked. They were first tested by colony PCR, after miniprep, they were examined by digestion check and send out for sequencing. All the tests confirmed that the construct was inserted into the pSB1C3 vector. | ||
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Latest revision as of 14:24, 26 September 2012
TuYV coat protein with part of the readthrough with His-tag
This part contains TuYV coat protein and part of readthrough with His-Tag on the N terminus. Because There are iGEN illegal sites in TuYV readthrough part, we just isolate part of the readthrough, which does not contain the illegal sites.
Turnip Yellows Virus (TuYV) is member of the genus Polerovirus and family Luteoviridae, which is positive sense RNA virus. The viral gene encoding the coat protein and part of the readthrough for TuYV, (GenBank: NC_003743.1 (3483..5495)) was obtained from a plasmid encoding the entire viral genome (GenBank: X13063.1). With designed primers, iGEM prefix and His-tag reporter protein was added to the 5 prime end and suffix were added 3 prime end of the this construct gene separately.
Then the PCR products were digested, ligated to pSB1C3 backbone, and transformed into DH5 alpha competent E.coli cells. Because we used white and red screening, the white colonies, which are colonies with right inserts, were checked. They were first tested by colony PCR, after miniprep, they were examined by digestion check and send out for sequencing. All the tests confirmed that the construct was inserted into the pSB1C3 vector.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 545
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
*Safety notice*
This part is isolated from a virus and, when assembled, will form particles that very much resemble this virus in size and shape. However, no additional viral information is stored in this part and viral infection and/or replication can therefore be ruled out. It is completely safe for use in normal laboratory circumstances.