A novel fluorescent timer based on bicistronic expression strategy in Caenorhabditis elegans.
Fluorescent timers are useful tools for studying the spatial and temporal cellular or molecular events. Based on the trans-splicing mechanism in Caenorhabditis elegans, we constructed a "fluorescent timer" through bicistronic expression of two fluorescent proteins with different maturation times. When used in vivo, this "timer" changes its color over time and therefore can be used to monitor the activity of the targeted promoters in C. elegans. Using this "timer", we have successfully traced the time-dependent activity of myo-3 promoter which drives expression in body wall muscle and vulval muscle. We found that the myo-3 promoter started to be active about 7 h after egg-laying and sustained its activity in the following hatching process. We have also determined the myo-3 promoter activity during larval development by this "timer". We anticipate that more new "fluorescent timers" with variable time-resolution could be designed by bicistronic expression of different fluorescent protein pairs.
Chen
Mao-Rong
MR
Key Laboratory of Molecular Biophysics, Ministry of Education, and Institute of Biophysics & Biochemistry, Huazhong University of Science and Technology, Wuhan 430074, PR China.
Yang
Song
S
Niu
Wei-pin
WP
Li
Zhao-Yu
ZY
Meng
Ling-Feng
LF
Wu
Zheng-Xing
ZX
D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans / Q000196:embryology* / Q000235:genetics*; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005453:Fluorescence; D018507:Gene Expression Regulation, Developmental; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics; D009132:Muscles / Q000196:embryology* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020038:RNA, Spliced Leader; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics; D020040:Trans-Splicing
D008164:Luminescent Proteins; D020038:RNA, Spliced Leader; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins