Construction of a T7 phage display library of human tongue squamous carcinoma

WANG Jing-tao; et al

doi:10.13406/j.cnki.cyxb.2010.11.044

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Construction of a T7 phage display library of human tongue squamous carcinoma

更新时间：2024-01-17

- Construction of a T7 phage display library of human tongue squamous carcinoma
- Journal of Chongqing Medical University Vol. 35, Issue 11, Pages: 1687-1689(2010)
- 作者机构：
  
  重庆医科大学附属口腔医院口腔颌面外科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2010.11.044
  CLC： R739.8
- Published：2010，
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[1]王景涛,季平,邓诚,伍明毅,刘凯,孙守娟,邹波.利用T7噬菌体展示技术构建人舌鳞癌细胞多肽文库[J].重庆医科大学学报,2010,35(11):1687-1689.

WANG Jing-tao, et al. Construction of a T7 phage display library of human tongue squamous carcinoma. [J]. Journal of Chongqing Medical University 35(11):1687-1689(2010)
[1]王景涛,季平,邓诚,伍明毅,刘凯,孙守娟,邹波.利用T7噬菌体展示技术构建人舌鳞癌细胞多肽文库[J].重庆医科大学学报,2010,35(11):1687-1689. DOI： 10.13406/j.cnki.cyxb.2010.11.044.

WANG Jing-tao, et al. Construction of a T7 phage display library of human tongue squamous carcinoma. [J]. Journal of Chongqing Medical University 35(11):1687-1689(2010) DOI： 10.13406/j.cnki.cyxb.2010.11.044.

摘要

目的:本研究拟构建人舌鳞癌细胞株Tca8113的T7噬菌体展示多肽文库

为进一步筛选和鉴定与舌鳞癌相关的蛋白或多肽打下基础。方法:人舌癌细胞株Tca8113

QiaGen法提取细胞mRNA

逆转录合成双链cDNA后与T7Select 10-3载体连接

体外包装并扩增得到T7噬菌体展示多肽文库。结果:构建了人舌鳞癌细胞株Tca8113的T7噬菌体展示多肽文库

原始文库重组滴度为鳞癌1.7×106 pfu

扩增后滴度为3×1010 pfu/mL。随机从文库中抽取10个克隆PCR扩增插入片断

电泳结果显示构建的文库中均有插入片段

文库插入率高达100%。所构建的文库插入片断大小在（0.4～1.0）kbp之间。结论:成功构建了人舌癌细胞株Tca8113的T7噬菌体展示多肽文库

为下一步筛选和鉴定与舌癌相关的蛋白或多肽奠定了基础。

Abstract

Objective:To construction human tongue squamous carcinoma T7 phage display peptides library from the cell of human tongue squamous carcinoma cell line Tca8113.Methods:mRNA was isolated from the cells of human tongue squamous carcinoma cell line Tca8113 with the methods of QiaGen

and then mRNA was reversely transcribed into double stranded cDNA.After digestion

the cDNA was inserted into T7 Select 10-3 vector.The phage display peptides library was constructed by package reaction in vitro and plate proliferation.Plaque assay and PCR were used to evaluate the library.Results:T7 phage display peptides library was established.Plaque assay showed the titer of the library was 1.7×106.The phage titer of the amplified library was 3×1010 pfu/mL.PCR amplification of random plaque showed insert ratio of 100% in the library.With insert range from 0.4 kbp to 1.0 kbp.Conclusion:The phage display peptides library from human tongue squamous carcinoma cell line Tca8113was constructed.

关键词

噬菌体展示多肽库人舌鳞癌TCA8113

Keywords

Phage displayPeptides libraryHuman tongue squamous carcinomaTCA8113

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