Regulation and expression of bacteria type pepc2 in Chlamydomonas reinhardtiiand and its effect on protein and lipid content
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College of Fisheries and Life Sciences,Shanghai Ocean University,College of Fisheries and Life Sciences,Shanghai Ocean University,College of Fisheries and Life Sciences,Shanghai Ocean University,College of Fisheries and Life Sciences,Shanghai Ocean University,Institute of Plant Physiology and Ecology,Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences,College of Fisheries and Life Sciences,Shanghai Ocean University,College of Fisheries and Life Sciences,Shanghai Ocean University

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Q785

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    Abstract:

    Phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) plays an important role in plant carbon metabolism, which can affect protein and lipid metabolism in higher plant, while studies on green alga PEPC are limited. Chlamydomonas reinhardtii is the model organism in green alga, and its chloroplast transformation is mature and efficient.In order to get high expression quality of protein and lipid content, chloroplast transformation method could be applied. In this study, the sequence with 639 bp (about 1/7 length) of C. reinhardtii CS I was cloned, then C. reinhardtii was transformed by the empty plasmid pASapI, vectors pASapI-reve-Crpepc2 and pASapI-forw-Crpepc2, to generate “blank,” “forward,” and“reverse” mutants by particle bombardment, respectively.The relative expression of Crpepc2 were detected with RT-qPCR, it showed that Crpepc2 expression of reverse mutant decreased by 89.97%, and that of forward mutant increased by 201.16%. Mean while the contents of protein and lipid were detected, the data showed that total protein of forward mutant rose by 37.03%, and the lipid content of reverse mutant enhanced by 70.32%, moreover, fatty acid of reverse mutant increased obviously. In conclusion, bacteria type PEPC in Chlamydomonas reinhardtii not only can improve protein and lipid content, but also can make contribution to foreign protein expression and high lipid content engineered microalgae.

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冯思豫,施定基,贾晓会,朱嘉诚,米华玲,贾睿,何培民.莱茵衣藻“细菌型”pepc2基因叶绿体表达及其对油脂蛋白质含量的影响[J].上海海洋大学学报,2017,26(5):641-650.
FENG Siyu, SHI Dingji, JIA Xiaohui, ZHU Jiacheng, MI Hualing, JIA Rui, HE Peimin. Regulation and expression of bacteria type pepc2 in Chlamydomonas reinhardtiiand and its effect on protein and lipid content[J]. Journal of Shanghai Ocean University,2017,26(5):641-650.

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History
  • Received:December 05,2016
  • Revised:May 12,2017
  • Adopted:June 15,2017
  • Online: September 25,2017
  • Published:
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