[Construction of retroviral vector with human brain-derived neurotrophic factor gene expression and in the fibroblasts expression].
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.


CONCLUSIONS
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.
Yang
Cheng
C
Stomatological Center, General Hospital of Chinese People's Armed Police Forces, Beijing, 100039, P.R. China.
Yan
Rongzeng
R
Guo
Hongyan
H
Gu
Xiaoming
X
D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics*; D002478:Cells, Cultured; D005347:Fibroblasts; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D016716:PC12 Cells; D051381:Rats; D017207:Rats, Sprague-Dawley; D012190:Retroviridae / Q000235:genetics; D014162:Transfection
D019208:Brain-Derived Neurotrophic Factor