Construction of Lentiviral Expression Vector Carrying SOX9 Gene and Its Stable Expression in LNcap Cell

Wang Jiang; Duan Jun Jun; Zhang Xiao Yu; Jia Xiao; Wang Xiao Bo; He Ying Hong

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Wang Jiang , Duan Jun Jun , Zhang Xiao Yu , Jia Xiao , Wang Xiao Bo , He Ying Hong . Construction of Lentiviral Expression Vector Carrying SOX9 Gene and Its Stable Expression in LNcap Cell[J]. Journal of Kunming Medical University, 2018, 39(02): 5-9.

Citation:

Wang Jiang , Duan Jun Jun , Zhang Xiao Yu , Jia Xiao , Wang Xiao Bo , He Ying Hong . Construction of Lentiviral Expression Vector Carrying SOX9 Gene and Its Stable Expression in LNcap Cell[J]. Journal of Kunming Medical University, 2018, 39(02): 5-9.

Citation:

Wang Jiang , Duan Jun Jun , Zhang Xiao Yu , Jia Xiao , Wang Xiao Bo , He Ying Hong . Construction of Lentiviral Expression Vector Carrying SOX9 Gene and Its Stable Expression in LNcap Cell[J]. Journal of Kunming Medical University, 2018, 39(02): 5-9.

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Construction of Lentiviral Expression Vector Carrying SOX9 Gene and Its Stable Expression in LNcap Cell

Wang Jiang ^1,2,
Duan Jun Jun ^1,2,
Zhang Xiao Yu ^1,2,
Jia Xiao ^1,2,
Wang Xiao Bo ^1,2,
He Ying Hong ^1,2

Funds:

基金：国家自然科学基金资助项目 (31360288);

Received Date: 2017-10-23

Abstract

Abstract

Objective To construct the lentiviral expression vector of SOX9 gene and establish a LNcap cell strain with stable expression of SOX9. Me thods SOX9 gene was amplified by PCR and cloned into lentiviral expression vector p LVX-IRES-Puro. p LVX-IRES-FLAG-SOX9 recombinant plasmid was verified by restriction enzyme digestion and DNA sequencing. Then we gained recombinant virus particles in packaging cell HEK 293 and infected LNcap cell. The monoclonal LNcap cell strain stably expressed were obtained through puromycin screening.The m RNA level and protein level in the infected LNcap cell were detected by q RT-PCR and Western blot respectively. Re s ults Restriction enzyme digestion and sequencing demonstrated that SOX9 c DNA was successfully cloned into p LVX-IRES-FLAG lentiviral vector. After the transfection to LNCa P cells, the monoclonal cell strain of stably expressed SOX9 were obtained by puromycin screening, which showed expression of SOX9 m RNA detected by q RT-PCR. Western blot analysis revealed that SOX9 protein expressed markedly. Conclus ion The recombinant lentiviral vector bearing human SOX9 c DNA has been successfully constructed, and the exogenous expression of SOX9 in LNcap cells was achieved.
- SOX9,
- Lentiviral vector,
- LNcap cell,
- Liposome tansfection

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References(15)

References

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