Construction of HPV16 E6 siRNA expressing recombinants and identification of transfection efficiency in vitro

CHENG Haien; et al

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Construction of HPV16 E6 siRNA expressing recombinants and identification of transfection efficiency in vitro

更新时间：2024-01-17

- Construction of HPV16 E6 siRNA expressing recombinants and identification of transfection efficiency in vitro
- Journal of Chongqing Medical University Issue 5, Pages: 636-640(2006)
- 作者机构：
  
  1. 重庆医科大学基础医学院生物化学与分子生物学教研室
  2. 重庆医科大学公共卫生学院生殖医学教研室
- 作者简介：
- 基金信息：
- DOI：
  CLC： R737.33
- Published：2006，
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[1]成海恩,张溢,张翠莉,石华,潘巍巍,易发平,马永平,宋方洲.表达HPV-16 E6基因siRNA真核载体的构建及体外转染效率鉴定[J].重庆医科大学学报,2006(05):636-640+676.

CHENG Haien, et al. Construction of HPV16 E6 siRNA expressing recombinants and identification of transfection efficiency in vitro. [J]. Journal of Chongqing Medical University (5):636-640(2006)
[1]成海恩,张溢,张翠莉,石华,潘巍巍,易发平,马永平,宋方洲.表达HPV-16 E6基因siRNA真核载体的构建及体外转染效率鉴定[J].重庆医科大学学报,2006(05):636-640+676. DOI：

CHENG Haien, et al. Construction of HPV16 E6 siRNA expressing recombinants and identification of transfection efficiency in vitro. [J]. Journal of Chongqing Medical University (5):636-640(2006) DOI：

摘要

目的:采用RNA干扰技术选择性的沉默宫颈癌Ca Ski细胞株中HPV-16 E6基因的表达。为观察HPV-16 E6基因的小干扰片段能否在Ca Ski细胞特异性的抑制E6基因的表达

构建能在哺乳动物细胞中表达E6小干扰RNA的真核表达质粒

并观察其转染效率。方法:根据HPV-16 E6基因序列

设计特异性的小干扰片段

并将其定向克隆到带有卡那霉素抗性和增强绿色荧光蛋白的真核表达载体pGenesil-1上

将其转染至Ca Ski细胞中

观察荧光表达

鉴定其转染效率。结果:通过酶切鉴定和序列测定

成功的构建二条表达小干扰RNA的质粒及其阴性对照质粒

并成功的转染到Ca Ski细胞株中

转染效率达到50%以上。结论:siRNA真核表达载体的构建及体外转染的成功为下一步研究奠定了良好的基础。

Abstract

Objective:To silence the expression of HPV16 E6 gene in Ca Ski cell line with RNA interference technique.In order to understand if the HPV16 E6 siRNA could inhibit the expression of E6 gene specifically

recombinant eukaryotic vectors were constructed and transfected into Ca Ski cell line.The transfection efficiency was observed.Methods: According to the sequence of HPV16 E6 gene

the E6 siRNA was designed and cloned into the EGFP reporter plasmid pGenesil-1

and then transfected into CaSki cell line.The fluorescence expression was observed and the transfection efficiency was identified.Results: Two siRNA expressing recombinants and the corresponding negative control vector were constructed and transfected into Ca Ski cell successfully

and the transfection efficiency achieved about 50%.Conclusion: The construction of recombinant E6 siRNA eukaryotic vectors and transfection of them into Ca Ski cell line successfully established a favourable foundation for further study.

关键词

HPV16E6基因RNA干扰转染

Keywords

HPV16 E6 geneRNA interferencetransfection

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