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	Establishment of a hek293t cell line able to site-specifically integrate and stably express gdnf by raav-2 vector
	Zhang, Jinju; Zhang, Yun; Liu, Xiaomei; Xiang, Jingjing; Zhang, Chun1
刊名	Electronic journal of biotechnology
	2016-07-01
卷号	22页码:75-80
关键词	Glial cell line-derived neurotrophic factor Multiplicity of infection Recombinant adeno-associated virus 2 Site-specific integration Transduction
ISSN号	0717-3458
DOI	10.1016/j.ejbt.2016.05.001
通讯作者	Zhang, chun(chunzhdr@163.com)
英文摘要	Background: using recombinant adeno-associated virus 2 (raav-2), we attempted to establish a hek293t cell line that is able to site-specifically integrate and stably express glial cell line-derived neurotrophic factor (gdnf). results: recombinant vector with enhanced green fluorescent protein (egfp) and gdnf (ptr-p5-egfp-ires-gdnf), as well as that carrying rep genes and sv40 promoters (psvav2) were constructed and packed. hek293t cells were co-infected with raav-2/egfp-gdnf and raav-2/svav2 virus separately at 1 x 10(4), 1 x 10(5), and 1 x 10(6) of multiplicity of infection (moi). the efficiency of transduction was detected using flow cytometry. additionally, the infected hek293t cells were separately validated by touchdown polymerase chain reaction (pcr) and western-blot. after 72 h of transduction, the rate of egfp positive cell was 22%, 45% and 49% at the mois of 1 x 10(4), 1 x 10(5) and 1 x 10(6), respectively. on the 3rd, 6th and 9th day of cell passage, there was no significant difference in the cell viability and proliferation rate between transduction and control groups. importantly, touchdown pcr showed that there was a specific pcr amplified product band in the lane of infected cells. furthermore, gdnf expression was detected in the infected cells after 15 and 180 d of cultivation. conclusions: a hek293t cell line able to site-specifically integrate and stably express gdnf was established. (c) 2016 pontificia universidad catolica de valparaiso. production and hosting by elsevier b.v. all rights reserved.
WOS关键词	MEDIATED GENE-TRANSFER ; NEUROTROPHIC FACTOR ; NIGROSTRIATAL SYSTEM ; DOPAMINERGIC-NEURONS ; LENTIVIRUS VECTORS ; THERAPY ; MODEL ; AAV ; REGENERATION ; DISEASES
WOS研究方向	Biotechnology & Applied Microbiology
WOS类目	Biotechnology & Applied Microbiology
语种	英语
出版者	UNIV CATOLICA DE VALPARAISO
WOS记录号	WOS:000380541700011
内容类型	期刊论文
URI标识	http://www.corc.org.cn/handle/1471x/2374477
专题	中国科学院大学
通讯作者	Zhang, Chun
作者单位	1.Chinese Acad Sci, Suzhou Inst Biomed Engn & Technol, CAS Key Lab Biomed Diagnost, Suzhou 215163, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Zhang, Jinju,Zhang, Yun,Liu, Xiaomei,et al. Establishment of a hek293t cell line able to site-specifically integrate and stably express gdnf by raav-2 vector[J]. Electronic journal of biotechnology,2016,22:75-80.
APA	Zhang, Jinju,Zhang, Yun,Liu, Xiaomei,Xiang, Jingjing,&Zhang, Chun.(2016).Establishment of a hek293t cell line able to site-specifically integrate and stably express gdnf by raav-2 vector.Electronic journal of biotechnology,22,75-80.
MLA	Zhang, Jinju,et al."Establishment of a hek293t cell line able to site-specifically integrate and stably express gdnf by raav-2 vector".Electronic journal of biotechnology 22(2016):75-80.

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