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[The wild-type alpha-synuclein over-expression to induce the protein aberrant aggregation of alpha-synuclein in HEK293 cells in vitro].
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2006 Feb; 23(1):19-22.ZY

Abstract

OBJECTIVE

To investigate over-expression of wild-type alpha-synuclein inducing the aberrant aggregation of alpha-synuclein in HEK293 cell in vitro.

METHODS

The cDNA encoding the human alpha-synuclein without the stop code was cloned into PGEM T-easy vector. Using enzyme map and DNA sequencing analyzed and determined the recombinant plasmid, and then sub-clone the alpha-synuclein cDNA fragment into pEGFP-N1 vector. The recombinant plasmids alpha-synuclein-pEGFP were transfected into HEK293 cells by lipofectamin 2000. The aberrant aggregation of alpha-synuclein was measured by EGFP fluorescence, anti-alpha-synuclein immunocytochemistry. The inclusions in the cultured cells were identified with HE staining.

RESULTS

The restriction enzyme map suggested that eukaryotic expression vector for human wild-type alpha-synuclein gene was constructed successfully. By EGFP fluorescence, anti-alpha-synuclein immunocytochemistry, it could be observed that the alpha-synuclein protein could aggregate in cytoplasm and the Lewy body-like inclusions found in cytoplasm of cultured cells.

CONCLUSION

The over-expression of wild-type alpha-synuclein can induce protein aberrant aggregation and Lewy body-like inclusions formation in cytoplasm of HEK293 cell in vitro.

Authors+Show Affiliations

Chen T
Department of Neurology, Xiangya Hospital, Central South University, Changsha, Hunan 410008, P.R.China.
Tang BS
No affiliation info available
Liao XP
No affiliation info available
Yan XX
No affiliation info available
Zhang RX
No affiliation info available
Zhang YH
No affiliation info available
Tang JG
No affiliation info available
Cao L
No affiliation info available
Guo JF
No affiliation info available
Li J
No affiliation info available

MeSH

Cells, CulturedGene ExpressionHumansImmunohistochemistryInclusion BodiesLewy BodiesParkinson Diseasealpha-Synuclein

Pub Type(s)

English Abstract
Journal Article

Language

chi

PubMed ID

16456779

Citation

Chen, Tao, et al. "[The Wild-type Alpha-synuclein Over-expression to Induce the Protein Aberrant Aggregation of Alpha-synuclein in HEK293 Cells in Vitro]." Zhonghua Yi Xue Yi Chuan Xue Za Zhi = Zhonghua Yixue Yichuanxue Zazhi = Chinese Journal of Medical Genetics, vol. 23, no. 1, 2006, pp. 19-22.
Chen T, Tang BS, Liao XP, et al. [The wild-type alpha-synuclein over-expression to induce the protein aberrant aggregation of alpha-synuclein in HEK293 cells in vitro]. Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2006;23(1):19-22.
Chen, T., Tang, B. S., Liao, X. P., Yan, X. X., Zhang, R. X., Zhang, Y. H., Tang, J. G., Cao, L., Guo, J. F., & Li, J. (2006). [The wild-type alpha-synuclein over-expression to induce the protein aberrant aggregation of alpha-synuclein in HEK293 cells in vitro]. Zhonghua Yi Xue Yi Chuan Xue Za Zhi = Zhonghua Yixue Yichuanxue Zazhi = Chinese Journal of Medical Genetics, 23(1), 19-22.
Chen T, et al. [The Wild-type Alpha-synuclein Over-expression to Induce the Protein Aberrant Aggregation of Alpha-synuclein in HEK293 Cells in Vitro]. Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2006;23(1):19-22. PubMed PMID: 16456779.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [The wild-type alpha-synuclein over-expression to induce the protein aberrant aggregation of alpha-synuclein in HEK293 cells in vitro]. AU - Chen,Tao, AU - Tang,Bei-sha, AU - Liao,Xiao-ping, AU - Yan,Xin-xiang, AU - Zhang,Ru-xu, AU - Zhang,Yu-hu, AU - Tang,Jian-guang, AU - Cao,Li, AU - Guo,Ji-feng, AU - Li,Jing, PY - 2006/2/4/pubmed PY - 2006/4/11/medline PY - 2006/2/4/entrez SP - 19 EP - 22 JF - Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese journal of medical genetics JO - Zhonghua Yi Xue Yi Chuan Xue Za Zhi VL - 23 IS - 1 N2 - OBJECTIVE: To investigate over-expression of wild-type alpha-synuclein inducing the aberrant aggregation of alpha-synuclein in HEK293 cell in vitro. METHODS: The cDNA encoding the human alpha-synuclein without the stop code was cloned into PGEM T-easy vector. Using enzyme map and DNA sequencing analyzed and determined the recombinant plasmid, and then sub-clone the alpha-synuclein cDNA fragment into pEGFP-N1 vector. The recombinant plasmids alpha-synuclein-pEGFP were transfected into HEK293 cells by lipofectamin 2000. The aberrant aggregation of alpha-synuclein was measured by EGFP fluorescence, anti-alpha-synuclein immunocytochemistry. The inclusions in the cultured cells were identified with HE staining. RESULTS: The restriction enzyme map suggested that eukaryotic expression vector for human wild-type alpha-synuclein gene was constructed successfully. By EGFP fluorescence, anti-alpha-synuclein immunocytochemistry, it could be observed that the alpha-synuclein protein could aggregate in cytoplasm and the Lewy body-like inclusions found in cytoplasm of cultured cells. CONCLUSION: The over-expression of wild-type alpha-synuclein can induce protein aberrant aggregation and Lewy body-like inclusions formation in cytoplasm of HEK293 cell in vitro. SN - 1003-9406 UR - https://www.unboundmedicine.com/medline/citation/16456779/[The_wild_type_alpha_synuclein_over_expression_to_induce_the_protein_aberrant_aggregation_of_alpha_synuclein_in_HEK293_cells_in_vitro]_ DB - PRIME DP - Unbound Medicine ER -