Construction of RPS14 recombinant lentiviral vector and its expression in MUTZ-8 cells

罗静; 蒋文; 刘林; 王利

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Construction of RPS14 recombinant lentiviral vector and its expression in MUTZ-8 cells

更新时间：2026-01-09

- Construction of RPS14 recombinant lentiviral vector and its expression in MUTZ-8 cells
- Journal of Army Medical University Vol. 33, Issue 21, Pages: 2252-2255(2011)
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  CLC： R394-33;R394.3;R552.02
- Published：2011
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罗静,蒋文,刘林,王利.RPS14基因重组慢病毒载体的构建及其在MUTZ-8细胞中的表达[J].陆军军医大学学报(原第三军医大学学报),2011,33(21):2252-2255.

　Luo Jing,Jiang Wen,Liu Lin,Wang Li.Construction of RPS14 recombinant lentiviral vector and its expression in MUTZ-8 cells[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(21):2252-2255.
罗静,蒋文,刘林,王利.RPS14基因重组慢病毒载体的构建及其在MUTZ-8细胞中的表达[J].陆军军医大学学报(原第三军医大学学报),2011,33(21):2252-2255. DOI：

　Luo Jing,Jiang Wen,Liu Lin,Wang Li.Construction of RPS14 recombinant lentiviral vector and its expression in MUTZ-8 cells[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(21):2252-2255. DOI：

摘要

目的构建携带人RPS14基因重组慢病毒载体获得稳定产毒的细胞，并观察其在人MDS细胞株MUTZ-8细胞中的表达。方法通过反转录方法，获得目的基因RPS14；将目的基因克隆入慢病毒表达质粒pGC-FU Vector，构建含有RPS14基因的重组慢病毒载体pGC-FU RPS14，经PCR、酶切及测序鉴定其正确性；在Lipofectamine 2000介导下，将成功构建的质粒pGC-FU RPS14与慢病毒包装质粒pHelper 1.0、pHelper 2.0共转染293T细胞，通过实时定量PCR法测定病毒滴度，并将获得的重组慢病毒载体pGC-FU RPS14转染人MDS细胞株MUTZ-8，用流式细胞仪检测转染效率，RT-PCR检测靶细胞内RPS14 mRNA的表达。结果含有RPS14基因的重组慢病毒载体经PCR、酶切和测序鉴定构建正确

且测定的病毒滴度为2.0×10

TU/ml。流式细胞仪检测其转染效率为68.41%，RT-PCR检测到转染后RPS14 mRNA在靶细胞中的稳定表达。结论构建了携带人RPS14 基因的慢病毒载体，转染人MUTZ-8细胞株后能够稳定表达RPS14基因。

Abstract

Objective To construct a recombinant lentiviral vector carrying RPS14 gene

establish a stable packaging 293T cell lines

and observe RPS14 expression in human MDS cell line MUTZ-8. Methods RPS14 was obtained using reverse transcription polymerase chain reaction (RT-PCR). Then the RPS14 gene was cloned into a lentiviral vector pGC-FU to construct a recombinant lentiviral vector carrying RPS14 gene named pGC-FU RPS14

which was confirmed by PCR

restriction enzyme digestion and DNA sequencing. With the help of Lipofectamine 2000

293T cells were cotransfected with pGC-FU RPS14

pHelper 1.0 and pHelper 2.0. The titer of the virus was examined by real-time PCR. pGC-FU RPS14 was used to transfect human MDS cell line MUTZ-8. The transfection efficiency and RPS14 mRNA expression were detected by flow cytometry and RT-PCR

respectively. Results The recombinant lentiviral vector pGC-FU RPS14 was confirmed to be correctly constructed by PCR

restriction enzyme digestion and DNA sequencing; the assayed titer of virus was 2.0×10

Tu/ml. The transfection efficiency was 68.41%

and stable expression of RPS14 mRNA in MUTZ-8 cells was detected by RT-PCR. Conclusion A lentiviral vector carrying RPS14 gene is successfully constructed

which can deliver RPS14 gene into human MDS cell line MUTZ-8 for RPS14 stable expression.

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