Green fluorescent protein of the jellyfish <i>Aequorea macrodactyla</i>

LUO Wen-xin; ZHANG Jun; YANG Hai-jie; XIE Xiao-yan; QIN Ying-xue; PANG Shu-qiang; LI Shao-jin; XIA Ning-shao

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Article Navigation > Haiyang Xuebao > 2002 > 24(4): 82-91

LUO Wen-xin, ZHANG Jun, YANG Hai-jie, XIE Xiao-yan, QIN Ying-xue, PANG Shu-qiang, LI Shao-jin, XIA Ning-shao. Green fluorescent protein of the jellyfish Aequorea macrodactyla[J]. Haiyang Xuebao, 2002, 24(4): 82-91.

Citation:

LUO Wen-xin, ZHANG Jun, YANG Hai-jie, XIE Xiao-yan, QIN Ying-xue, PANG Shu-qiang, LI Shao-jin, XIA Ning-shao. Green fluorescent protein of the jellyfish Aequorea macrodactyla[J]. Haiyang Xuebao, 2002, 24(4): 82-91.

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Green fluorescent protein of the jellyfish Aequorea macrodactyla

Ministry of Education, Cell Biology and Tumor Cell Engineering Laboratory, Xiamen University, Xiamen 361005, China

Received Date: 2001-02-26
Rev Recd Date: 2001-04-26

Abstract

Abstract

A new green fluor escent protein gene-gfpxm was isolated from jellyfish Aequorea macrodactyla in the coastal region of East China Sea by the method of combination of PCR amplification and southern hybridization analysis.The gfpxm gene contains three extrons and two introns spread over 1042 bp of genomic DNA,the entire coding region of cDNAis 717 bp,being identical to the wild-type gfp-Aeqgfp 10 cDNA in length.The amino acid sequence of GFPxm deduced from the nucleotide sequence is 238-aa-residue,sharing homology of 83.6% with that of AeqGFP10.The entire coding sequence was cloned into the pTO-T7 expression vector and expressed in Ecoli. The expression yield of GFPxm was amounted to 50% of the total protein.Compared with GFP of Avictoria,the expressed GFPxm exhibited an excitation peak at a higher wave length of 476nm and an emission peak at a lower wave length 496 nm with a higher quantum yield of 10.The fluorescence of GFPxm is significant stable,showing strong resistant to heat,alkaline,denaturants and salts.
- Aequorea macrodactyla,
- green fluorescent protein gene,
- cloning and expression,
- fluorescent property

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References(21)

References

WARD W W, CODY C W, HART R C, et al. Sepectrophoton etric identity of the energy transfer chromophores in Renilla and Aequorea green fluorescent proteins[J]. Photobiochem Photobio, 1980, 31:611-615.

KAIN S R, ADAMS M, KONDEPUDI A, et al. Green fluorescent protein as a reporter of gene expression and protein localization[J]. Biotechniques, 1995, 19: 650-655.

WANG S, HAZELRIGG T. Implications for bcd mRNA localization from spatial distribution of in drosophila oogendsis[J].Nature, 1994, 369: 400-403.

BARTHMAIER P, FYRBERY E. Rapid communication monitoring development and pathology of drosophila indirect flight muscles using green fluorescent protein[J]. Dev Biol, 1995, 169: 770-774.

SCALES S J, PEPPERKOK R, KREIS T E. Visualization of ER-to-Golgi transport in living cells reveals a sequential mode of action for COPII and COPI[J]. Cell, 1997, 90:1 137-1 148.

ULLMAN E F, MARSHALL W F, SEDAT J W, et al. Mitosis in living budding yeast:anaphase A but no metaphase plate[J]. Science, 1999, 277: 574-578.

HU W, CHENG C L. Expression of Aequorea green fluorescent protein in plant cells[J]. FEBS Letters, 1995, 369: 331-334.

MOSS J B, ROSENTHAL N. Analysis of gene expression patterns in the embryonic mouse myotome with the green fluorescent protein[J]. J Cell Biochem, 1994, 180: 489-495.

许振祖,张金标.奥东一闽南近海的浮游水螅水母类,管水母类和钵水母类[J]厦门大学学报(自然科学版),1978,17(4):32

萨姆布鲁克J,弗里奇EF,曼尼阿蒂斯T.分子克隆操作指南(第二版)[M].金冬雁,黎孟枫等译北京:科学出版社,1992.

陈国珍,黄贤智,许金钩,等.荧光分析法(第二版)[M]北京:科学出版社,1990.16-17

PRASHER D C, ECKENRODE V K, WARD W W, et al. Primary structure of the Aequorea victoria green-fluorescent protein[J]. Gene, 1992, 111: 229-233.

罗文新,张军,夏宁邵,等一种带增强子的原核表达载体的构建及初步应用[J].生物工程学报,2000,16(5):578-581.

MORISE J G, SHIMOMURA O, JOHNSON F H, et al. Intermolecular energy transfer in the bioluminescent system of Aequorea[J]. Biochemistry, 13(194): 2 656～2 662.

WARD W W. Properties of the coelenterate green-fluorescent proteins[A]. Deluca M, Mcelroy W D. Bioluminescence and chemiluminescence:basic chemistry and analytical application[M]. New York: Academic Press Inc, 1981. 235-242.

WARD W W, BOKMAN S H. Reversible denaturation of aequorea green fluorescent protein: physical separation and characterization of the renatured protein[J]. Biochemistry, 1982, 21:4 535-4 540.

CHALFIE M, TU Y, EUSKIRCHEN G, et al. Green fluorescent protein as a marker for gene expression[J]. Science,1994, 263: 802-808.

HEIM R, CUBITT A, TSIEN R Y. Improved green fluorescence[J]. Nature, 1995, 373: 663-664.

HEIM R, PRASHER D C, TSIEN R Y. Wavelength mutations and posttrans-lational autoxidation of green fluorescent protein[J]. Proc Natl Acad Sci USA, 1994, 91:12 501-12 504.

HEIM R, TSIEN R Y. Engineering green fluorescent protein for improved brightness, longer wavelengths and fluorescence resonance energy transfer[J]. Curr Biol, 1996, 6: 178-182.

MIRO R D, SILVA C M, YOUVAN D C. Fluorescence resonance energy transfer between blueemitting and red-shifted excitation derivatives of the green fluorescent protein[J]. Gene, 1996, 173: 13-17.

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