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Journal of Zhejiang University SCIENCE B 2005 Vol.6 No.1 P.11-13

http://doi.org/10.1631/jzus.2005.B0011


Construction of a eukaryotic expression plasmid of Humanin


Author(s):  LUO Ben-yan, CHEN Xiang-ming, TANG Min, CHEN Feng, CHEN Zhi

Affiliation(s):  Department of Neurology, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, China; more

Corresponding email(s):   lobenyan@263.net

Key Words:  Humanin, Alzheimer's disease, Eukaryotic expression


LUO Ben-yan, CHEN Xiang-ming, TANG Min, CHEN Feng, CHEN Zhi. Construction of a eukaryotic expression plasmid of Humanin[J]. Journal of Zhejiang University Science B, 2005, 6(1): 11-13.

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author="LUO Ben-yan, CHEN Xiang-ming, TANG Min, CHEN Feng, CHEN Zhi",
journal="Journal of Zhejiang University Science B",
volume="6",
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publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.2005.B0011"
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%J Journal of Zhejiang University SCIENCE B
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%DOI 10.1631/jzus.2005.B0011

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T1 - Construction of a eukaryotic expression plasmid of Humanin
A1 - LUO Ben-yan
A1 - CHEN Xiang-ming
A1 - TANG Min
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A1 - CHEN Zhi
J0 - Journal of Zhejiang University Science B
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IS - 1
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%@ 1673-1581
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PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.2005.B0011


Abstract: 
Objective: To construct a eukaryotic expression plasmid pcDNA3.1(-)-humanin. Methods: The recombinant plasmid pGEMEX-1-humanin was digested with restriction endonucleases BamH I and Hind III and the humanin gene fragments, about 100 bp length, were obtained. Then the humanin gene fragments were inserted into eukaryotic expression vector pcDNA3.1(-) and the recombinant plasmids pcDNA3.1(-)-humanin were identified by sequencing. Results: Recombinant plasmid DNA successfully produced a band which had the same size as that of the humanin positive control. The sequence of recombinant plasmids accorded with the Humnain gene sequence. Conclusions: A eukaryotic expression plasmid of humanin was successfully constructed.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1] Hashimoto, Y., Ito, Y., Niikura, T., Shao, Z., Hata, M., Oyama, F., Nishimoto, I., 2001a. Mechanisms of neuroprotection by a novel rescue factor Humanin from Swedish mutant amyloid precursor protein. Biochem Biophys Res Commun, 283(2):460-468.

[2] Hashimoto, Y., Niikura, T., Ito, Y., Sudo, H., Hata, M., Arakawa, E., Abe, Y., Kita, Y., Nishimoto, I., 2001b. Detailed characterization of neuroprotection by a rescue factor HN against various Alzheimer’s disease-relevant insults. J Neurosci, 21(23):9235-9245.

[3] Hashimoto, Y., Niikura, T., Tajima, H., Yasukawa, T., Sudo, H., Ito, Y., Kita, Y., Kawasumi, M., Kouyama, K., Doyu, M., Sobue, G., Koide, T., Tsuji, S., Lang, J., Kurokawa, K., Nishimoto, I., 2001c. A rescue factor abolishing neuronal cell death by a wide spectrum of familial Alzheimer’s disease genes and Abeta. Proc Natl Acad Sci USA, 98(11):6336-6341.

[4] Luo, B.Y., Yuan, M., Tang, M., Chen, Z., 2004. Cloning and expression of the anti-AD protective peptide Humanin. Chin J Neurology, 37(3):231-233 (in Chinese).

[5] Niikura, T., Hashimoto, Y., Tajima, H., Nishimoto, I., 2002. Death and survival of neuronal cells exposed to Alzheimer’s insults. J Neurosci Res, 70(3):380-391.

[6] Terashita, K., Hashimoto, Y., Niikura, T., Tajima, H., Yamagishi, Y., Ishizaka, M., Kawasumi, M., Chiba, T., Kanekura, K., Yamada, M., Nawa, M., Kita, Y., Aiso, S., Nishimoto, I., 2003. Two serine residues distinctly regulate the rescue function of Humanin, an inhibiting factor of Alzheimer’s disease-related neurotoxicity: functional potentiation by isomerization and dimerization. J Neurochem, 85(6):1521-1538.

[7] Zou, P., Ding, Y., Sha, Y., Hu, B., Nie, S., 2003. Humanin peptides block calcium influx of rat hippocampal neurons by altering fibrogenesis of Abeta (1-40). Peptides, 24(5):679-685.

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