Yuan, GH; Xu, XY; Zhang, W; Zhang, WL; Cui, YL
; Qin, S; Liu, TZ发表期刊FRONTIERS IN MICROBIOLOGY
ISSN1664-302X
2019-08-02
卷号10页码:1749
关键词alpha tubulinbiolistic transformationflanking sequenceHaematococcus pluvialisselection marker
研究领域Microbiology
DOI10.3389/fmicb.2019.01749
产权排序[Yuan, Guanhua; Xu, Xiaoying; Zhang, Wei; Zhang, Wenlei; Liu, Tianzhong] Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Key Lab Biofuels, Qingdao, Shandong, Peoples R China; [Yuan, Guanhua; Zhang, Wei; Zhang, Wenlei; Liu, Tianzhong] Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Shandong Prov Key Lab Energy Genet, Qingdao, Shandong, Peoples R China; [Yuan, Guanhua] Univ Chinese Acad Sci, Beijing, Peoples R China; [Xu, Xiaoying] Yantai Marine Econ Res Inst, Yantai, Peoples R China; [Cui, Yulin; Qin, Song] Chinese Acad Sci, Yantai Inst Coastal Zone Res, Key Lab Coastal Biol & Biol Resource Utilizat, Yantai, Peoples R China
通讯作者Zhang, Wei(zhangwei@qibebt.ac.cn); Liu, Tianzhong(liutz@qibebt.ac.cn)
作者部门海岸带生物学与生物资源保护实验室
英文摘要Haematococcus pluvialis has high commercial value, yet it displays low development of genetic transformation systems. In this research, the endogenous 5' and 3' flanking sequences of the constitutive alpha tubulin (tub) gene were cloned along with its encoding region in H. pluvialis, in which some putative promoter elements and polyadenylation signals were identified, respectively. Three selection markers of tub/aadA, tub/hyr and tub/ble with three different antibiotic-resistance genes fused between the endogenous tub promoter (Ptub) and terminator (Ttub) were constructed and utilized for biolistic transformation of H. pluvialis. Stable resistant colonies with introduced aadA genes were obtained after bombardments of either H. pluvialis NIES144 or SCCAP K0084 with the tub/aadA cassette, the efficiency of which could reach up to 3 x 10(-5) per mu g DNA through an established manipulation flow. Two key details, including the utilization of culture with motile flagellates dominant and controlled incubation of them on membrane filters during bombardments, were disclosed firstly. In obtained transformants, efficient integration and transcription of the foreign tub/aadA fragments could be identified through genome PCR examination and qPCR analysis, nonetheless with random style instead of homologous crossover in the H. pluvialis genome. The presented selection marker and optimized transforming procedures in this report would strengthen the platform for genetic manipulation and modification of H. pluvialis.
文章类型Article
资助机构2016 Sino-Thai Cooperation Project by the National Natural Science Foundation of ChinaNational Natural Science Foundation of China [51561145015]; Shandong Provincial Key Laboratory of Energy Genetics [SDKLEG201805]; Synthetic Biology Technology Innovation Center of Shandong Province [sdsynbio-2018-PY-08]
收录类别SCI
语种英语
关键词[WOS]NUCLEAR TRANSFORMATION; ASTAXANTHIN; EXPRESSION; PROMOTER; CHLOROPHYCEAE; PCR; RECOMBINATION; ELEMENTS; GROWTH
研究领域[WOS]Microbiology
WOS记录号WOS:000478564700001
引用统计被引频次：6[WOS][WOS记录][WOS相关记录]
文献类型期刊论文
条目标识符http://ir.yic.ac.cnhttp://ir.yic.ac.cn/handle/133337/24885
专题海岸带生物学与生物资源利用重点实验室_海岸带生物学与生物资源保护实验室
作者单位1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Key Lab Biofuels, Qingdao, Shandong, Peoples R China;
2.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Shandong Prov Key Lab Energy Genet, Qingdao, Shandong, Peoples R China;
3.Univ Chinese Acad Sci, Beijing, Peoples R China;
4.Yantai Marine Econ Res Inst, Yantai, Peoples R China;
5.Chinese Acad Sci, Yantai Inst Coastal Zone Res, Key Lab Coastal Biol & Biol Resource Utilizat, Yantai, Peoples R China
推荐引用方式
GB/T 7714Yuan, GH,Xu, XY,Zhang, W,et al. Biolistic Transformation of Haematococcus pluvialis With Constructs Based on the Flanking Sequences of Its Endogenous Alpha Tubulin Gene[J]. FRONTIERS IN MICROBIOLOGY,2019,10:1749.
APAYuan, GH.,Xu, XY.,Zhang, W.,Zhang, WL.,Cui, YL.,...&Liu, TZ.(2019).Biolistic Transformation of Haematococcus pluvialis With Constructs Based on the Flanking Sequences of Its Endogenous Alpha Tubulin Gene.FRONTIERS IN MICROBIOLOGY,10,1749.
MLAYuan, GH,et al."Biolistic Transformation of Haematococcus pluvialis With Constructs Based on the Flanking Sequences of Its Endogenous Alpha Tubulin Gene".FRONTIERS IN MICROBIOLOGY 10(2019):1749.
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