BBa_K1634007 1 pmyo2-ChR2 pmyo2-ChR2-YFP 2015-09-13T11:00:00Z 2015-09-27T05:37:11Z 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_ 0 24889 24399 9 In stock true 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 component2453847 1 BBa_K1634003 component2453845 1 BBa_K1634002 annotation2453847 1 BBa_K1634003 range2453847 1 701 2509 annotation2453845 1 BBa_K1634002 range2453845 1 1 694 BBa_K1634003 1 BBa_K1634003 ChR2-YFP ( a kind of channelrhodopsin ) 2015-09-12T11:00:00Z 2015-09-13T11:52:49Z 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. 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. And if the cell is a muscle cell or a neuron, this tiny change may trigger a great change in the organism's activity. To make it easier to confirm the location and expression level of the ChR2, we construct a fusion protein of ChR2 and YFP. 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 ChR2-YFP, avoiding using the restriction site exits in it. false Xinhong Chen annotation2453249 1 ChR2-YFP range2453249 1 1 1809 BBa_K1634002 1 BBa_K1634002 pmyo2 ( a promoter in C.elegans ) 2015-09-12T11:00:00Z 2015-09-14T07:53:57Z We get Pmyo2 from the C.elegans' genomic DNA by PCR. Pmyo2 is a promoter which can drive the expression in the muscle of C.elegans especially the muscle in the head and pharynx.And we link the Pmyo2 with some channelrhodopsin which can be activated or supressed by the light. 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 can use them 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. false false _2051_ 25024 24399 9 false Since we use the restriction enzyme site to do the ligation, we should pay attention to the sequence of Pmyo2, avoiding using the restriction site exits in it. false Xinhong Chen annotation2453209 1 pmyo2 range2453209 1 1 694 BBa_K1634002_sequence 1 tgctgaactttacaccccgaacagcaatgtgtgcttcagcctaaaaaaaagtaagtgtgttaatcagtgcccccgattcttcattttttgcccctctctcccgtttcgtcggcaaaagaagagaaaataaagataagtctcaagataggttggtaatcgctaaagtggttgtgtggataagagtagcaaaatggcaggaagagcactttgcgcgcacacactgtactcattgttctggataaaattctctcgttgtttgccgtcggatgtctgcctctctgcattgagccggcttcttcactatctttagttaacctaaaatgccgtttcttttctcgtatccccactatcccgttgaggttctctgctctcttcgctccctaccgccagcgagcaactatccgtgggggcgccttgctcggaagatgggggggaagaaagaagatttttgctatttgcacttgagaaagagacttttcctgcgtcgatggttagagaacagtgtgcagacacttttcagctacctagaattacaattggatatccccgcctcccaatccacccacccagggaaaaagaagggctcgccgaaaatcaaagttatctccaggctcgcgcatcccaccgagcggttgacttctctccaccacttttcattttaaccctcgatcgtcagacacagaaatggattacg BBa_K1634007_sequence 1 tgctgaactttacaccccgaacagcaatgtgtgcttcagcctaaaaaaaagtaagtgtgttaatcagtgcccccgattcttcattttttgcccctctctcccgtttcgtcggcaaaagaagagaaaataaagataagtctcaagataggttggtaatcgctaaagtggttgtgtggataagagtagcaaaatggcaggaagagcactttgcgcgcacacactgtactcattgttctggataaaattctctcgttgtttgccgtcggatgtctgcctctctgcattgagccggcttcttcactatctttagttaacctaaaatgccgtttcttttctcgtatccccactatcccgttgaggttctctgctctcttcgctccctaccgccagcgagcaactatccgtgggggcgccttgctcggaagatgggggggaagaaagaagatttttgctatttgcacttgagaaagagacttttcctgcgtcgatggttagagaacagtgtgcagacacttttcagctacctagaattacaattggatatccccgcctcccaatccacccacccagggaaaaagaagggctcgccgaaaatcaaagttatctccaggctcgcgcatcccaccgagcggttgacttctctccaccacttttcattttaaccctcgatcgtcagacacagaaatggattacgtactagatggattatggaggcgccctgagtgccgttgggcgcgagctgctatttgtaacgaacccagtagtcgtcaatggctctgtacttgtgcctgaggaccagtgttactgcgcgggctggattgagtcgcgtggcacaaacggtgcccaaacggcgtcgaacgtgctgcaatggcttgctgctggcttctccatcctactgcttatgttttacgcctaccaaacatggaagtcaacctgcggctgggaggagatctatgtgtgcgctatcgagatggtcaaggtgattctcgagttcttcttcgagtttaagaacccgtccatgctgtatctagccacaggccaccgcgtccagtggttgcgttacgccgagtggcttctcacctgcccggtcattctcattcgcctgtcaaacctgacgggcttgtccaacgactacagcaggcgcaccatgggtctgcttgtgtctgatattggcacaattgtgtggggcgccacttccgccatggccaccggatacgtcaaggtcatcttcttctgcctgggtctgtgttatggtgctaacacgttctttcacgctgccaaggcctacatcgagggttaccacaccgtgccgaagggccggtgtcgccaggtggtgactggcatggcttggctcttcttcgtatcatggggtatgttccccatcctgttcatcctcggccccgagggcttcggcgtcctgagcgtgtacggctccaccgtcggccacaccatcattgacctgatgtcgaagaactgctggggtctgctcggccactacctgcgcgtgctgatccacgagcatatcctcatccacggcgacattcgcaagaccaccaaattgaacattggtggcactgagattgaggtcgagacgctggtggaggacgaggccgaggctggcgcggtaccggtagaaaaaatgagtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatgggtaagtttaaacatatatatactaactaaccctgattatttaaattttcagccaacacttgtcactactttcggttatggtcttcaatgcttcgccagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtaagtttaaacagttcggtactaactaaccatacatatttaaattttcaggtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttgtaagtttaaacatgattttactaactaactaatctgatttaaattttcagaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtcctaccaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaatag BBa_K1634003_sequence 1 atggattatggaggcgccctgagtgccgttgggcgcgagctgctatttgtaacgaacccagtagtcgtcaatggctctgtacttgtgcctgaggaccagtgttactgcgcgggctggattgagtcgcgtggcacaaacggtgcccaaacggcgtcgaacgtgctgcaatggcttgctgctggcttctccatcctactgcttatgttttacgcctaccaaacatggaagtcaacctgcggctgggaggagatctatgtgtgcgctatcgagatggtcaaggtgattctcgagttcttcttcgagtttaagaacccgtccatgctgtatctagccacaggccaccgcgtccagtggttgcgttacgccgagtggcttctcacctgcccggtcattctcattcgcctgtcaaacctgacgggcttgtccaacgactacagcaggcgcaccatgggtctgcttgtgtctgatattggcacaattgtgtggggcgccacttccgccatggccaccggatacgtcaaggtcatcttcttctgcctgggtctgtgttatggtgctaacacgttctttcacgctgccaaggcctacatcgagggttaccacaccgtgccgaagggccggtgtcgccaggtggtgactggcatggcttggctcttcttcgtatcatggggtatgttccccatcctgttcatcctcggccccgagggcttcggcgtcctgagcgtgtacggctccaccgtcggccacaccatcattgacctgatgtcgaagaactgctggggtctgctcggccactacctgcgcgtgctgatccacgagcatatcctcatccacggcgacattcgcaagaccaccaaattgaacattggtggcactgagattgaggtcgagacgctggtggaggacgaggccgaggctggcgcggtaccggtagaaaaaatgagtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatgggtaagtttaaacatatatatactaactaaccctgattatttaaattttcagccaacacttgtcactactttcggttatggtcttcaatgcttcgccagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtaagtttaaacagttcggtactaactaaccatacatatttaaattttcaggtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttgtaagtttaaacatgattttactaactaactaatctgatttaaattttcagaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtcctaccaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaatag 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