Construction and expression of retroviral vector pLEGFP-N1-TERT in preparation of seed cells for skin tissue engineering.
Construction and expression of retroviral vector pLEGFP-N1-TERT in preparation of seed cells for skin tissue engineering.
复制标题
逆转录病毒载体pLEGFP-N1-TERT的构建及表达用于皮肤组织工程种子细胞的制备。
DOI：
10.1016/s1995-7645(13)60171-6
复制
发表时间：
2013
影响因子：
3.1
通讯作者：
Zhi
中科院分区：
文献类型：
--
作者：
Ting Tan;Zhi
ObjectiveTo construct the retroviral vector pLEGFP-N1-telomerase reverse transcriptase (TERT) and to investigate the expression of TERT in neonatal mouse hypodermal cells.MethodsThe polymerase chain reaction (PCR)-amplifiedTERTgene was inserted into plasmid pLEGFP-N1. The positive clone was identified by restriction enzyme digestion and sequencing, then was transfected into packaging cells to produce retrovirus particles. Neonatal mouse hypodermal cells were infected with the virus to generate a stable cell line. TheTERTmRNA expression level, telomerase activity, and enhanced green fluorescent protein (EGFP) expression level were analyzed.ResultsRetroviral vector pLEGFP-N1-TERT was constructed successfully, and a stable cell line of neonatal mouse hypodermal cells expressing EGFP was established. Western blot and immunohistochemical assay showed that the expression level of TERT was significantly elevated in the neonatal mouse hypodermal cells.ConclusionsA high titer of retrovirus pLEGFP-N1-TERT mediates high-level expression of the exogenousTERTgene in the neonatal mouse hypodermal cells. This protocol has potential applications for skin tissue engineering and cell transplantation therapy.
影响因子：
4.4
作者：
Robertson, DM;Li, L;Jester, JV
通讯作者：
Jester, JV
影响因子：
6.5
作者：
Brem, Harold;Kodra, Arber;Tomic-Canic, Marjana
通讯作者：
Tomic-Canic, Marjana