Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (41): 7701-7705.doi: 10.3969/j.issn.1673-8225.2011.41.025

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Rapid construction of herpes simplex virus type I vector for gene therapy 

Cai Xiao-hui, Zhang Qin-xian   

  1. Basic Medical College of Zhengzhou University, Zhengzhou   450052, Henan Province, China
  • Received:2011-07-14 Revised:2011-08-29 Online:2011-10-08 Published:2011-10-08
  • Contact: Zhang Qin-xian, Professor, Doctoral supervisor, Basic Medical College of Zhengzhou University, Zhengzhou 450052, Henan Province, China
  • About author:Cai Xiao-hui★, Studying for master’s degree, Basic Medical College of Zhengzhou University, Zhengzhou 450052, Henan Province, China huixiaocai@126.com

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Abstract:

BACKGROUND: Herpes simplex virus typeⅠ(HSV-1) vector has been currently widely used due to its unique advantages, but a rapid and effect method is needed to construct this vector.
OBJECTIVE: To develop a rapid method using Cre/loxp site specific recombination system to construct HSV-1 vector.
METHODS: HSV-1 was isolated and c66-SV40-cre plasmid containing Cre recombinase was used to transfect Vero cells. Then HSV-1 HSVLoxp was constructed. Shuttle vector pShuttle-SV40-Cre-Loxp-IRES and HSV-I vector HSV-GDNF were constructed. The positive strains screened by HAT culture medium were identified by PCR taking GDNF as primer. After amplification, titer was determined.
RESULTS AND CONCLUSION: A strain of HSV-1 was isolated and pHV-TK-GFP was constructed successfully. Recombinant virus HSVtk-Loxp-GFP01 without Us3 gene was isolated. HSV-1 HSVLoxp and shuttle vector pShuttle- SV40-Cre-Loxp-IRES were successfully constructed. GDNF gene was successfully obtained and transferred into HSV-1 vector. Thus, HSV-1 vector expressing GDNF was successfully constructed and the titer was 2.25×106 IU/mL.

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