Cloning and expression of the aequorin genes from jellyfish aequorea and characterization of aequorins activities

LUO Wen-xin; ZHANG Jun; LI Shao-wei; CHENG Tong; CHEN Min; LI Shao-jin; XIA Ning-shao

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Article Navigation > Haiyang Xuebao > 2004 > 26(4): 110-117

LUO Wen-xin, ZHANG Jun, LI Shao-wei, CHENG Tong, CHEN Min, LI Shao-jin, XIA Ning-shao. Cloning and expression of the aequorin genes from jellyfish aequorea and characterization of aequorins activities[J]. Haiyang Xuebao, 2004, 26(4): 110-117.

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Cloning and expression of the aequorin genes from jellyfish aequorea and characterization of aequorins activities

1.
Cell Biology and Tumor Cell Engineering Laboratory of Ministry of Education Xiamen University, Xiamen 361005, China;College of Oceanography and Environmental Sciences, Xiamen University, Xiamen 361005, China
2.
Cell Biology and Tumor Cell Engineering Laboratory of Ministry of Education Xiamen University, Xiamen 361005, China

Received Date: 2001-04-16
Rev Recd Date: 2001-08-13

Abstract

Abstract

Two new aequorin genes aeqxm and aeqxxm were isolated from fellyfish Aequorea macr odactyla and A.parva respectively,which are commonly found in the warmer waters on the coastal region of East China Sea.The DNA sequences of the two genes have no introns and each one contains an ORF of 585 bp in full-length encoding a 195-aa protein.The two genes of aeqxm and aeqxxm share nucleotide homologies of 80.7% and 85.1% with AEVAQ 440X respectively,and the corresponding proteins share amino acid homologies of 84.7% and 84.2% with AEVAQ 440X.High amino acid homology was found between apoaeqxm and apoaeqx xm.The two genes were cloned into expression vector pTO-T7 respectively,and the expression yields were amounted to 40% of the total protein in E.coli BL21.The activities of the two photoproteins were reconstituted by incubating the expressed apoproteins with coelenterazine f.In the presence of Caion,both of the regenerated aeqxm and aeqxxm exhibited an emission peak at the wave length of 470 nm.
- Aequorea macrodactyla,
- Aequoreaparva,
- aequorin,
- expression

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References(19)

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