Difference between revisions of "Part:BBa K1618028"

Latest revision as of 16:27, 21 September 2015

Chloroplast Transit Peptide

BBa_K1618028 contains a synthetic plastid transit peptide from the consensus of multiple RuBisCo small subunit genes. In an N-terminal Fusion, this peptide provides subcellular localisation to the chloroplast. It is assembled in a Golden Gate Universal Acceptor Plasmid (pUAP1) in the pSB1C3 backbone according to RFC106. It is a Golden Gate part in the plant standard syntax, not a BioBrick.

NRP-UEA 2015 team used this part in our plant constructs. Constructs K1618029-36 contained a 35s promoter, chloroplast transit peptide, one of four acyltransferases, and this new 35s terminator parts. In parts K1618029-32 fused the recombinant protein was fused to a fluorescent a reporter tag to confirm the localisation to the chloroplast (Figure 1). The parts were tested by transferring into Agrobacterium tumefaciens and infiltration into plants. The results confirm that the chloroplast transit peptide is functional and able to direct proteins to plant plastids.

Figure 1: Constructs BBa_K1618029-032 contain a yellow fluorescent protein, as well as a chloroplast transit peptide. These are confocal microscopy images of the constructs infiltrated into Nicotiana benthamiana, in which the red stains are the chloroplast, and the yellow fluorescence is construct (a) BBa_K1618029, (b) BBa_K1618031, (c) BBa_K1618032, and (d) BBa_K1618030

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 <partinfo>BBa_K1618028 short</partinfo>
-This is synthetic a 5'UTR, omega (Tobacco Mosaic Virus. The chloroplast transit peptide provides subcellular localisation to the chloroplast. It is assembled in a Golden Gate Universal Acceptor Plasmid (pUAP1) in the pSB1C3 backbone according to RFC106. It is a Golden Gate part in the plant standard syntax, not a BioBrick.
+BBa_K1618028 contains a synthetic plastid transit peptide from the consensus of multiple RuBisCo small subunit genes. In an N-terminal Fusion, this peptide provides subcellular localisation to the chloroplast. It is assembled in a Golden Gate Universal Acceptor Plasmid (pUAP1) in the pSB1C3 backbone according to RFC106. It is a Golden Gate part in the plant standard syntax, not a BioBrick.
-The Chloroplast Transit Peptide was used by NRP-UEA 2015 House of Carbs. It was used in 8 composite constructs in order to make sure that the parts reached the chloroplast, where starch is produced. For 4 of these constructs, a Yellow Fluorescent Protein tag was added, in order to visualise if the parts did in fact reach the chloroplast. Using a confocal microscope, they obtained the following images, in which the chloroplast is red and our protein with the fluorescent tag is yellow.
+NRP-UEA 2015 team used this part in our plant constructs. Constructs K1618029-36 contained a 35s promoter, chloroplast transit peptide, one of four acyltransferases, and this new 35s terminator parts. In parts K1618029-32 fused the recombinant protein was fused to a fluorescent a reporter tag to confirm the localisation to the chloroplast (Figure 1). The parts were tested by transferring into <i>Agrobacterium tumefaciens</i> and infiltration into plants. The results confirm  that the chloroplast transit peptide is functional and able to direct proteins to plant plastids.
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 ''Figure 1: Constructs BBa_K1618029-032 contain a yellow fluorescent protein, as well as a chloroplast transit peptide. These are confocal microscopy images of the constructs infiltrated into Nicotiana benthamiana, in which the red stains are the chloroplast, and the yellow fluorescence is construct (a) BBa_K1618029, (b) BBa_K1618031, (c) BBa_K1618032, and (d) BBa_K1618030''
-The results above suggest that the chloroplast transit peptide is functional in regards to the localisation of parts to the chloroplast.
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