| Along with the development of fossil fuels such as oil, coal, while bringing economic benefits and the following is serious pollution and energy shortage. So nowadays, the development of a new renewable energy has become the primary problem of the world. Biodiesel, as a kind of renewable energy gets the attention of the researchers all over the world, but its high production cost keeps people away. Because of its strong environmental adaptability and growth cycle is short, fatty, high photosynthetic efficiency microalage can be used as a raw material to product biodiesel, using geothermal water to culture microalgae, this not only reduces the production cost of biodiesel production, on the other hand also improved the environment, so the microalgae biodiesel is expected to become important renewable energy alternatives to fossil fuels.In order to improve the oil content of microalgae on the molecular level to build a kind of high oil microalgae, so total RNA of perilla frutescens is extracted by RNAiso Plus kit. It is used as a template for RT-PCR to clone cDNA coding region of DGAT1 gene of perilla frutescens. And cloning vector pMD-DGAT1 is constructed. Cloning vector pMD-DGAT1 and empty vector pBI121 are double-enzyme digested with Xba I and BamH I. Then expression vector pBI121-DGATl is constructed by linking the fragments. At last,1657 bp fragment is obtained by cloning and has 97% base similarity to DGAT1 gene of GenBank. The pBI121 - DGAT1 is successfully transformed into Scenedesmus quadricanda taking the use of electric conversion. Then using geothermal wastewater to cultivate the engineered microalgae with DGAT1, the optimum culture conditions is pH 7.5-8.5, the temperature is 25℃, light intensity 5500 lx, ventilation 220 L/h, incubation time 11d. The removal rate of COD, TN, TP and F- in the geothermal wastewater is 78%,91%,89% and 81%, and under the condition of the culture, the transformed Scenedesmus quadricanda approximately double the oil content as before. |
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