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  • Title: [Construction of retroviral vector with human brain-derived neurotrophic factor gene expression and in the fibroblasts expression].
    Author: Yang C, Yan R, Guo H, Gu X.
    Journal: Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi; 2009 May; 23(5):588-91. PubMed ID: 19514583.
    Abstract:
    OBJECTIVE: To construct human brain-derived neurotrophic factor retroviral vector-pLXSN (hBDNF-pLXSN), and to evaluate the bioactivity of hBDNF. METHODS: The genome mRNA was extracted from embryonic brain tissue of a 5-month-old infant, the hBDNF gene sequence was obtained with RT-PCR technology, and hBDNF-pLXSN constructed in vitro was used to infect the fibroblasts (NIH/3T3). The expression of hBDNF was identfied by the immunohistochemistry method, and the NIH/3T3 and BDNF biological activities were determined by culture of the PC12 cells and dorsal root ganglia. RESULTS: The hBDNF-pLXSN was constructed successfully by sequencing analyses. The infected NIH/3T3 showed positive expression of hBDNF. The infected NIH/3T3 could product hBDNF. Bioactivity of the products could support the PC12 cell survival and neurite growth in the primary cultures of dorsal root ganglia neurons of mice. CONCLUSION: hBDNF-pLXSN virus has the ability to infect NIH/3T3 and make it expressed and secreted hBDNF with the biological activity. It can be used to treat facial paralysis as a gene therapy.
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