BBa_K1634000 1 BBa_K1634000 Pmyo2-ChR2-YFP (C.elegans) 2015-09-06T11:00:00Z 2015-09-07T02:24:10Z We get Pmyo2 from the C.elegans' genomic DNA by PCR. ChR2 is a channelrhodopsin in green algae, but we didn't get it directly from the algae. Instead, we get ChR2 from some plamids by PCR. Then we ligated the Pmyo2 into PPD95.75, a vector commonly used in C.elegans. Finally, we construted the fused protein, ChR2-YFP,and ligated it into PPD95.75_Pmyo2. Pmyo2 is a promoter which can drive the expression in the muscle of C.elegans especially the muscle in the head and pharynx. ChR2 is a kind of channelrhodopsin. Channelrhodopsins are a subfamily of retinylidene proteins (rhodopsins) that function as light-gated ion channels.[1] They serve as sensory photoreceptors in unicellular green algae, controlling phototaxis: movement in response to light.[2] When we express the channelrhodopsins in some specific cells in organisms and shed specific light on them, we can activate or supress the specific ion channel to change the activity of the cell. According to this thought in optogenetic, we linked the Pmyo2，specific promoter in the muscle of head, with the ChR2, then we use the light source assembled by ourselves to shed light of specific wave on the C.elegans,changging the condition of the muscle. Since we have learned that the chosing of direction in C.elegans depends on the muscle in the head, we can observe the obvious change in moving pattern of the C.elegans after we shed the light. In other words,we try to construct a light-sensed locomotion controlling system in C.elegans. And we achieve that goal by using this part to control the movement of the head, the movement of the whole C.elegans. References: 1. Nagel G, Ollig D, Fuhrmann M, Kateriya S, Musti AM, Bamberg E, Hegemann P (June 2002). "Channelrhodopsin-1: a light-gated proton channel in green algae". Science 296 (5577): 2395???8. doi:10.1126/science.1072068 2. Sineshchekov OA, Jung KH, Spudich JL (June 2002). "Two rhodopsins mediate phototaxis to low- and high-intensity light in Chlamydomonas reinhardtii". Proc. Natl. Acad. Sci. U.S.A. 99 (13): 8689???94. doi:10.1073/pnas.122243399. false false _2051_ 24399 24399 9 false Since we use the restriction enzyme site to do the ligation, we should pay attention to the sequence of Pmyo2 and ChR2-YFP, avoiding using the restriction site exits in it. false Xinhong Chen annotation2445389 1 BamHI range2445389 1 695 700 annotation2445397 1 ChR2 range2445397 1 701 2509 annotation2445386 1 Pmyo2 range2445386 1 1 694 BBa_K1634000_sequence 1 tgctgaactttacaccccgaacagcaatgtgtgcttcagcctaaaaaaaagtaagtgtgttaatcagtgcccccgattcttcattttttgcccctctctcccgtttcgtcggcaaaagaagagaaaataaagataagtctcaagataggttggtaatcgctaaagtggttgtgtggataagagtagcaaaatggcaggaagagcactttgcgcgcacacactgtactcattgttctggataaaattctctcgttgtttgccgtcggatgtctgcctctctgcattgagccggcttcttcactatctttagttaacctaaaatgccgtttcttttctcgtatccccactatcccgttgaggttctctgctctcttcgctccctaccgccagcgagcaactatccgtgggggcgccttgctcggaagatgggggggaagaaagaagatttttgctatttgcacttgagaaagagacttttcctgcgtcgatggttagagaacagtgtgcagacacttttcagctacctagaattacaattggatatccccgcctcccaatccacccacccagggaaaaagaagggctcgccgaaaatcaaagttatctccaggctcgcgcatcccaccgagcggttgacttctctccaccacttttcattttaaccctcgatcgtcagacacagaaatggattacgggatccatggattatggaggcgccctgagtgccgttgggcgcgagctgctatttgtaacgaacccagtagtcgtcaatggctctgtacttgtgcctgaggaccagtgttactgcgcgggctggattgagtcgcgtggcacaaacggtgcccaaacggcgtcgaacgtgctgcaatggcttgctgctggcttctccatcctactgcttatgttttacgcctaccaaacatggaagtcaacctgcggctgggaggagatctatgtgtgcgctatcgagatggtcaaggtgattctcgagttcttcttcgagtttaagaacccgtccatgctgtatctagccacaggccaccgcgtccagtggttgcgttacgccgagtggcttctcacctgcccggtcattctcattcgcctgtcaaacctgacgggcttgtccaacgactacagcaggcgcaccatgggtctgcttgtgtctgatattggcacaattgtgtggggcgccacttccgccatggccaccggatacgtcaaggtcatcttcttctgcctgggtctgtgttatggtgctaacacgttctttcacgctgccaaggcctacatcgagggttaccacaccgtgccgaagggccggtgtcgccaggtggtgactggcatggcttggctcttcttcgtatcatggggtatgttccccatcctgttcatcctcggccccgagggcttcggcgtcctgagcgtgtacggctccaccgtcggccacaccatcattgacctgatgtcgaagaactgctggggtctgctcggccactacctgcgcgtgctgatccacgagcatatcctcatccacggcgacattcgcaagaccaccaaattgaacattggtggcactgagattgaggtcgagacgctggtggaggacgaggccgaggctggcgcggtaccggtagaaaaaatgagtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatgggtaagtttaaacatatatatactaactaaccctgattatttaaattttcagccaacacttgtcactactttcggttatggtcttcaatgcttcgccagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtaagtttaaacagttcggtactaactaaccatacatatttaaattttcaggtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttgtaagtttaaacatgattttactaactaactaatctgatttaaattttcagaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtcctaccaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaatag igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 Chris J. Myers James Alastair McLaughlin 2017-03-06T15:00:00.000Z