Liang Chuan-yu

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Co-Authors

Shixi Liu

West China Hospital of Sichuan University

Junming Xian

Sichuan University

Guangyao Zhou

Anhui University of Science and Technology

Yuquan Wei

Sichuan University

Jumei Zhao

Yan'an University

Ling Tian

South China Agricultural University

Tao Liu

Affiliated Hospital of Southwest Medical University

Cooperative Institutions

Sichuan University

State Key Laboratory of Biotherapy

West China Hospital of Sichuan University

China Agricultural University

Guangzhou University

University of Economics and Law

Henan University of Economic and Law

Fudan University

Science and Technology Department of Sichuan Province

University of Chinese Academy of Sciences

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[Construction of recombinant adenovirus vector carrying cell cycle controlling gene-p14ARF].

Junming Xian Guangyao Zhou Liang Chuan-yu Shixi Liu

OBJECTIVE: The recombinant adenovirus vector carrying p14ARF gene was constructed for using in the interference therapy in signal transduction of laryngeal squamous cell carcinoma. METHODS: The total cDNA fragment of p14ARF was cloned into the shuttle plasmid pAdTrack-CMV, with the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the E.Coli BJ5183 and the recombinant adenoviral plasmid was generated. The adenoviruses were packaged and amplified in the 293 cells. Then the viral titer was checked by GFP. RESULTS: The recombinant adenovirus vector carrying p14ARF was constructed successfully. The viral titer was 2.3 x 10(9). CONCLUSION: The recombinant adenovirus vector could introduce p14ARF gene into the laryngeal squamous cell carcinoma line or tumor tissue effectively, which would provide experimental basis for the mechanisms and further study of the interference therapy in signal transduction of laryngeal squamous cell carcinoma.

Transduction (biophysics)

Shuttle vector

p14arf

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[Cloning human heat shock protein 90beta-cDNA and constructing its eukaryon vector].

Sheng wu yi xue gong cheng xue za zhi = Journal of biomedical engineering = Shengwu yixue gongchengxue zazhi (2006)

Tao Liu Jumei Zhao Ling Tian Yuquan Wei Liang Chuan-yu

Cloning (programming)

Trizol

Agarose

Cloning vector

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[Construction of the recombinant adenovirus vector with EGFR antisense cDNA of controlling cell cycle].

PubMed (2006)

Junming Xian Guangyao Zhou Liang Chuan-yu Shixi Liu

To recombine the adenovirus vector carrying EGFR sence/antisense cDNA which takes part in control of cell cycle.The 1032 bp EGFR sence/antisense cDNA fragment was cloned into the shuttle plasmid pAdTrack-CMV. The resultant plasmid and the backbone plasmid pAdEasy-1 were transferred into E. coli BJ5183 for homologous recombination, and the recombinant adenoviruses were generated in cells. The recombinant adenoviruses were packaged and amplified in the 293 cells. Then the viral titer was detected by GFP.The recombinant adenovirus vector carrying EGFR sence/antisense cDNA to control the cell cycle was constructed successfully. The viral titers were 2.2 x 10(9) efu/mL and 2.5 X 10(9) efu/mL respectively.The recombinant adenovirus vector constructed by us could introduce EGFR antisense cDNA into the laryngeal squamous cell carcinoma line or tumor tissue, which would provide an experiment basis to study further the interfered mechanism of signal transdution and the therapies of laryngeal squamous cell carcinoma.

Shuttle vector

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Construction of pcDNA3.1(+)/hHSP90β plasmid and its expression in eukaryote cells

Chinese Journal of Otorhinolaryngology-skull Base Surgery (2005)

Liang Chuan-yu

Objective To establish a construct that can express human HSP90β in eukaryote cells in vitro.Methods Total RNA was isolated from human NPC and HSP90βcDNA was prepared by RT-PCR.The purified HSP90β fragment was cloned into PGEMT Easy T Vector,and then sub-cloned into pcDNA3.1(+) vector.The resulted pcDNA3.1(+)/hHSP90β plasmid was verified by restriction enzyme digestion and DNA sequencing.COS cells were transfected with this plasmid and the expression of HSP90β was examined by RT-PCR and Western blot analysis.Results Sequencing and restriction enzyme digestion analysis showed HSP90β cDNA was correctly cloned into vector pcDNA3.1(+).HSP90β was expressed in COS cells transfected with pcDNA3.1(+)/hHSP90β plasmid.Conclusion The constructed eukaryote expression plasmid pcDNA3.1(+)/hHSP90β could express HSP90β in eukaryote cells in vitro.

Eukaryote

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