Application of chlamydomonas reinhardtii VMPL1 gene to regulation of cadmium tolerance of chlamydomonas reinhardtii
A rhinestone and tolerance technology, applied in the fields of biology and sewage treatment, can solve the problems of complex operation, high cost, secondary pollution, etc., and achieve the effect of mature technology system and simple method
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[0036] 1. Obtaining Chlamydomonas reinhardtii transformants by electroporation
[0037] (1) Prepare the aphⅧ fragment
[0038] PJMG-aphⅧ plasmid containing aphⅧ fragment (Zhangfeng Hu, Yinwen Liang, Wei He, Junmin Pan * , Cilia Disassembly with Two Distinct Phases of Regulation, CellReports, 2015, 10(11): 1803-1810) was digested with EcoRI enzyme at 37°C overnight, and the aphⅧ fragment was separated by 1% agarose gel electrophoresis (2118bp, sequence such as SEQ ID NO :3), the result of electrophoresis after enzyme digestion is as follows figure 1 As shown, M is DL2000 DNA Marker, 1 is the pJMG-aphⅧ plasmid EcoRI digestion product, and the band around 2k is the aphⅧ fragment. The aphⅧ fragment was recovered from the gel with Sangon’s Gel Recovery Kit (Cat. No. B518191-0100), and the concentration of the recovered DNA fragment was measured with a Q5000 ultra-micro nucleic acid detector, and stored at -20°C for future use.
[0039] (2) Chlamydomonas cells prepared for electr...
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