Advance Search
Ma Qiong, Xiang Jian-Wei, Mou Ruo-Lan, Deng La-Qing, Wang Yun, Zhou Ming. Expression and photochemical properties of cyanobacteriochrome Alr1966GAF2 and its mutants[J]. Plant Science Journal, 2020, 38(4): 551-557. DOI: 10.11913/PSJ.2095-0837.2020.40551
Citation: Ma Qiong, Xiang Jian-Wei, Mou Ruo-Lan, Deng La-Qing, Wang Yun, Zhou Ming. Expression and photochemical properties of cyanobacteriochrome Alr1966GAF2 and its mutants[J]. Plant Science Journal, 2020, 38(4): 551-557. DOI: 10.11913/PSJ.2095-0837.2020.40551
shu

Expression and photochemical properties of cyanobacteriochrome Alr1966GAF2 and its mutants

  • 1. Key Laboratory of Biologic Resources Protection and Utilization of Hubei Province, Hubei University for Nationalities, Enshi, Hubei 445000, China;

  • 2. College of Biological Science and Technology, Hubei University for Nationalities, Enshi, Hubei 445000, China;

  • 3. Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China

Funds: 

This work was supported by grants from the Project of Science and Technology of Enshi (D20190025), National Natural Science Foundation of China (21472055), Incubation Project for High-Level Scientific Research Achievements of Hubei Minzu University(PY20020), and Project of Student Research Training from Hubei Minzu University (X201910517274, X201910517231).

More Information
  • Received Date: October 23, 2019
  • Revised Date: December 25, 2019
  • Available Online: October 31, 2022
  • Published Date: August 27, 2020
    Graphical Abstract
    • Abstract
  • To construct pET-alr1966gaf2, a fragment of alr1966gaf2 was amplified by polymerase chain reaction (PCR) from Anabaena sp. PCC7120, and then inserted into pET-30a(+). For over-expression, pET-alr1966gaf2 was transformed into Escherichia coli BL21(DE3) containing pACYC-ho1-pcyA and biliproteins were co-expressed successfully. Results showed that bili-Alr1966GAF2 had a sequential reversible photoconversion in three different states. We also detected red fluorescence reversible photoconversion of Alr1966GAF2 in 15E-P514 nm/15Z-P428 nm forms. Via site-directed mutagenesis, we mutated conserved Cys into Ala in the conserved DXCF motif of Alr1966GAF2, resulting in Alr1966GAF2(C72A). Alr1966GAF2(C72A)-PCB showed strong red fluorescence and high fluorescence quantum yield of 0.11. Furthermore, Alr1966GAF2(C72A) could bind to phycoerythrobilin (PCB) and biliverdin (BV) covalently, with strong red fluorescence. Therefore, as a red fluorescent protein, Alr1966GAF2(C72A) could be further developed as a fluorescent probe and applied in life sciences.
    • FullText(HTML)
    • References (16)
  • [1]
    Sun YF, Xu JG, Tang K, Miao D, Gärtner W, et al. Orange fluorescent proteins constructed from cyanobacteriochromes chromophorylated with phycoerythrobilin[J]. Photochem Photobiol, 2014, 13(5):757-763.
    [2]
    Fushimi K, Enomoto G, Ikeuchi M, Narikawa R. Distinctive properties of dark reversion kinetics between two red/green-type cyanobacteriochromes and their application in the photoregulation of cAMP synthesis[J]. Photochem Photobiol, 2017, 93(3):681-691.
    [3]
    Ma Q, Zheng XJ, Zhou Z, Zhou M. Fluorescence spectra properties of All4261 binding with phycocyanobilin in E. coli[J]. J Appl Spectrosc, 2014, 81(3):437-441.
    [4]
    Sato M, Ikeuchi M. A biliverdin-binding cyanobacteriochrome from the chlorophyll d-bearing cyanobacterium Acaryochloris marina[J]. Sci Rep, 2015, 7950(5):1-10.
    [5]
    Fushimi K, Nakajima T, Aono Y, Yamamoto T, Win NN, et al. Photoconversion and fluorescence properties of a red/green-type cyanobacteriochromeAM1_C0023g2 that binds not only phycocyanobilin but also biliverdin[J]. Front Microbiol, 2016, 26(7):1-12.
    [6]
    Rumyantsev KA, Shcherbakova DM, Zakharova NI, Verkhusha VV, Turoverov KK. Design of near-Infrared single-domain fluorescent protein GAF-FP based on bacterial phytochrome[J]. Cell Tissue Biol, 2017, 11(1):16-26.
    [7]
    Richie CT, Whitaker LR, Whitaker KW, Necarsulmer J, Baldwin HA, et al. Near-infrared fluorescent protein iRFP713 as a reporter protein for optogenetic vectors, a transgenic cre-reporter rat, and other neuronal studies[J]. J Neurosci Meth, 2017, 284:1-14.
    [8]
    Tang K, Ding WL, Höppner A, Zhao C, Zhang L, et al. The terminal phycobilisome emitter, LCM:A light-harvesting pigment with a phytochrome chromophore[J]. Proc Natl Acad Sci USA, 2015, 112(52):15880-15885.
    [9]
    Zhang J, Wu XJ, Wang ZB, Chen Y, Wang X, Zhou M, et al. Fused-gene approach to photoswitchable and fluorescent bili-proteins[J]. Angew Chem Int Edit, 2010, 49(32):5456-5458.
    [10]
    Reichhart E, Ingles-Prieto A, Tichy AM, McKenzie C, Janovjak H, et al. A phytochrome sensory domain permits receptor activation by red light[J]. Angew Chem Int Edit, 2016, 55(21):6339-6342.
    [11]
    Bugaj LJ, Choksi AT, Mesuda CK, Kane RS, Schaffer DV. Optogenetic protein clustering and signaling activation in mammalian cells[J]. Nat methods, 2013, 10(3):249-253.
    [12]
    王晓辉. 增强型绿色荧光蛋白在DH5α大肠杆菌中的表达[J]. 食品科学, 2010, 31(21):200-203.

    Wang XH. Expression of enhanced green fluorescent protein in DH5α[J]. Food science, 2011, 31(21):200-203.
    [13]
    Leopold AV, Chernov KG, Shemetov AA, Verkhusha VV. Neurotrophin receptor tyrosine kinases regulated with near-infrared light[J]. Nat Commun, 2019, 10:1129.
    [14]
    Bugaj LJ, Spelke DP, Mesuda CK. Regulation of endogenous transmembrane receptors through optogenetic Cry2 clustering[J]. Nat Commun, 2015, 6:6898.
    [15]
    Xu ZQ, Han JX, Tang QY, Ding WL, Miao D, et al. Far-red light photoacclimation:Chromophorylation of FR induced α- and β-subunits of allophycocyanin[J]. Biochimica et Biophysica Acta, 2016, 1857(9):1607-1616.
    [16]
    马涛, 姚明月, 刘延琳. 酿酒酵母APAl的定点突变以及与增强型绿色荧光蛋白EGFP基因的共表达[J]. 食品科学, 2015, 36(23):200-203.

    Ma T, Yao MY, Liu YL. Site-directed mutagenesis of Saccharomyces cerevisiae APA1 and co-expression with EGFP[J]. Food science, 2015, 36(23):200-203.
    • Related Articles

  • Related Articles

    [1]Kang Hongwei, Yue Kangjie, Liu Huixin, Wang Jiali, Tian Xuping. Effect of sex and leaf shape on gas exchange parameters and chlorophyll fluorescence characteristics in Sabina chinensis (L.) Ant[J]. Plant Science Journal, 2024, 42(6): 791-799. DOI: 10.11913/PSJ.2095-0837.23391
    [2]Han Jia-Xin, Zheng Hao, Zhang Qiong, Zhong Cai-Hong. Research advances in the metabolism and regulation of carbohydrate in fruit trees[J]. Plant Science Journal, 2020, 38(1): 143-149. DOI: 10.11913/PSJ.2095-0837.2020.10143
    [3]Zu Kui-Ling, Dong Shu-Bin, Li Jian-Xia, Xu Shen-Jian, Zhao Liang-Cheng. Differentially expressed genes analysis of terpenoid biosynthesis related to aril development in Celastrus orbiculatus Thunb.[J]. Plant Science Journal, 2017, 35(2): 276-282. DOI: 10.11913/PSJ.2095-0837.2017.20276
    [4]LIANG Jia-Wen, LIU Ai-Jie, MA Bing-Xin, WANG You-Wei. Simultaneous Determination of Six Flavonoid Compounds in Lotus Leaves by High Performance Liquid Chromatography[J]. Plant Science Journal, 2015, 33(6): 861-866. DOI: 10.11913/PSJ.2095-0837.2015.60861
    [5]CHANG Jing-Ling, DENG Xiao-Li, ZHANG Jun-Xia, ZHAO Xu-Na, YANG Jian-Lei. GC-MS Analysis of Linolenic Acid and Linoleic Acid in Chinese Trichosanthes kirilowii Oil[J]. Plant Science Journal, 2009, 27(5): 564-568.
    [6]LI Miao, LOU Yi-Ceng, YANG Han, HU Jia-Xing, SU Feng-Ping. Study on the Fingerprint Chromatogram of the Compound Giant Knotweed Rhizome by High Performance Liquid Chromatography[J]. Plant Science Journal, 2009, 27(4): 446-450.
    [7]HE Ya-Ting, LI Ming, LIU Wen-Zhi, ZHANG Quan-Fa, DANG Gao-Di. Comparison of Gas Exchange Traits of 30 Plant Species in Subalpine Meadow in Foping National Reserve of Qinling Mountains[J]. Plant Science Journal, 2007, 25(5): 451-456.
    [8]MA Jin-E, JIN Ze-Xin, LI Jun-Min. Analysis of Flavonoids in Endangered Plant Sinocalycanthus chinensis Using Thin Layer Chromatography[J]. Plant Science Journal, 2007, 25(4): 366-370.
    [9]LEI An-Ping, HU Zhang-Li, WONG Yuk-Shan, TAM Fung-Yee. Bioconcentration and Metabolism of Polycyclic Aromatic Hydrocarbons (PAHs) by Algae[J]. Plant Science Journal, 2005, 23(3): 291-298.
    [10]Mei Xinguo. A LOW-PRESSURE CHROMATOGRAPHIC APPARATUS AND ITS USE IN PHYTOCHEMISTRY[J]. Plant Science Journal, 1986, 4(3): 297-302.

Catalog

    Get Citation
    PDF
    XML
    Article views (562) PDF downloads (804) Cited by()
    Turn off MathJax
    Article Contents
    Abstract
    References
    Related Articles
    Copyright © 2022 Plant Science Journal 鄂ICP备05004779号-3
    Address：No. 201, Jiufeng 1st Road, Donghu High tech Zone, WuhanPostal Code：430074

    Supported by: Beijing Renhe Information Technology Co., Ltd.  Baidu Analytics

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return