Difference between revisions of "Part:BBa K883500"
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This plasmid contains the TuYV coat protein, which can be used as building block for TuYV VLPs. | This plasmid contains the TuYV coat protein, which can be used as building block for TuYV VLPs. | ||
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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 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 suffix were added to the 5 prime end and 3 prime end of the coat protein 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 TuYV coat protein was inserted into the pSB1C3 vector. | ||
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Revision as of 14:13, 26 September 2012
TuYV coat potein
This plasmid contains the TuYV coat protein, which can be used as building block for TuYV VLPs.
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 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 suffix were added to the 5 prime end and 3 prime end of the coat protein 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 TuYV coat protein 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 524
- 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.