Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (37): 6922-6926.doi: 10.3969/j.issn.1673-8225.2010.37.019

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Construction and identification of a recombinant adenovirus vector containing human heat shock protein 70 gene  

Han Shi-wei, Zhang Zhong-tao, Wang Yu   

  1. Department of General Surgery, Beijing Friendship Hospital, Capital University of Medical Sciences, Beijing  100050, China
  • Online:2010-09-10 Published:2010-09-10
  • Contact: Zhang Zhong-tao, Doctor, Professor, Doctoral supervisor, Chief physician, Department of General Surgery, Beijing Friendship Hospital, Capital University of Medical Sciences, Beijing 100050, China zhangzht@medmail.com.cn
  • About author:Han Shi-wei☆, Studying for doctorate, Department of General Surgery, Beijing Friendship Hospital, Capital University of Medical Sciences, Beijing 100050, China cco1985@yahoo.com.cn

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Abstract:

BACKGROUND: Adenovirus vectors have been widely used as a low high effectiveness and low toxicity gene vector, however, little is known about a recombinant adenovirus vector containing human heat shock protein 70 (Hsp70) gene.
OBJECTIVE: To construct a recombinant adenovirus vector containing human Hsp70 gene and to express the gene efficiently in eukaryotic cells.
METHODS: The recombinant adenovirus vector carrying Hsp70 gene was constructed with AdMax system and then transferred human embryonic kidney 293 (HEK293) cells to produce recombinant adenovirus. The exogenous gene expression was detected by Western blot and the viral titer was tested.
RESULTS AND CONCLUSION: The significant cytopathic effects were observed in transfected HEK293 cells, and then the recombinant virus was harvested and purified. The expression of green fluorescence protein could be observed by fluorescence microscope and the expression of Hsp70 could be detected by Western blot. The viral titer was 1×1011 efu/mL. Recombinant adenovirus vector was constructed successfully and packed in HEK293 cells.

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