JIA Qing-hua, et al. Construction and identification of recombinant adenovirus vector of keratinocyte growth factor. [J]. Journal of Chongqing Medical University 36(1):64-67(2011)
JIA Qing-hua, et al. Construction and identification of recombinant adenovirus vector of keratinocyte growth factor. [J]. Journal of Chongqing Medical University 36(1):64-67(2011) DOI: 10.13406/j.cnki.cyxb.2011.01.002.
Construction and identification of recombinant adenovirus vector of keratinocyte growth factor
Objective:To construct and identify a recombinant adenovirus vector expressing keratinocyte growth factor(KGF) gene.Methods:KGF gene was amplified from the plasmid pIRES2-EGFP-KGF by polymerase chain reaction(PCR) and then inserted into the plasmid pShuttle-CMV to construct pShuttle-KGF.After being confirmed by restriction enzyme digestion and DNA sequencing
the DNA encoding KGF in the new structure was inserted into the vector of recombinant plasmid adenovirus and confirmed by restriction enzyme digestion.Then the human embryonic kidney cell line 293 was transfected with correctly identified pAdxsi-KGF
and the recombinant adenovirus were amplified and purified
and the virus titer was detected using 50% tissue culture infective dose(TCID50) assay.The expression of the green fluorescent protein and KGF were detected by fluorescent microscope and ELISA after the recombinant adenovirus transfected human lung cancer A549 cell line.Results:The recombinant adenovirus pAdxsi-KGF was constructed and amplified with titer of 1.6×1010 pfu/ml;the green fluorescence proteins could be observed under fluorescent microscope in A549 24 h after transfection and with a stronger degree after 48 h
and KGF expression was detected by ELISA.Conclusion:The recombinant adenovirus pAdxsi-KGF was constructed successfully
which will be a good foundation for the research on the therapy of KGF for pulmonary fibrosis.
Cloning and expression analysis of surfactant-associated protein D in cotton rat with lung injury induced by lipopolysaccharide
Construction and expression of recombinant GRP78-expressed adenovirus and GRP78 expression in HepG2
Construction and expression of a recombinant hepatits c virus gene vector
Cloning and expressing of vacA gene related segment with vacuolating activity of Helicobacter pylori
Cloning and expression of survivin gene in eukaryotic cells
Related Author
Feng Yi
Ren Luo
Huang Daochao
Li Yasha
Zhong Haiying
Zhao Li
Wu Shuang
Zhang Xiang
Related Institution
Department of Pediatric Research Institute,the Children’s Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders,Chongqing Key Laboratory of Pediatrics,Chongqing International Science and Technology Cooperation Center for Child Development and Disorders
Department of Clinical Laboratory,People’s Hospital of Yubei District of Chongqing
Key Laboratory of Molecular Biology on Infectious Diseases,Ministry of Education,Chongqing Medical University
Department of Laboratory,Chongqing Medical University