Construction of eukaryotic expression system of a cell surface molecule——human CD40 ligand

田坤; 钱桂生

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Construction of eukaryotic expression system of a cell surface molecule——human CD40 ligand

更新时间：2026-01-09

- Construction of eukaryotic expression system of a cell surface molecule——human CD40 ligand
- Journal of Army Medical University Vol. 29, Issue 10, Pages: 963-966(2007)
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  CLC： R392-33;R394.2;R730.51
- Published：2007
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田坤,钱桂生.人CD40L真核表达体系的建立[J].陆军军医大学学报(原第三军医大学学报),2007,29(10):963-966.

　TIAN Kun,QIAN Gui-sheng.Construction of eukaryotic expression system of a cell surface molecule——human CD40 ligand[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(10):963-966.
田坤,钱桂生.人CD40L真核表达体系的建立[J].陆军军医大学学报(原第三军医大学学报),2007,29(10):963-966. DOI：

　TIAN Kun,QIAN Gui-sheng.Construction of eukaryotic expression system of a cell surface molecule——human CD40 ligand[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(10):963-966. DOI：

摘要

目的构建人CD40L真核表达体系并鉴定其在转染细胞中的表达。方法以DNA重组技术构建CD40L的真核表达载体pcDNA3.1(+)-CD40L，脂质体法转染于人小细胞肺癌细胞株h446，流式细胞技术鉴定其在转染细胞中的表达。结果双酶切及DNA测序证明，CD40L DNA插入表达载体pcDNA3.1(+) 序列及方向正确，转染肺小细胞肺癌细胞株h446后，经流式细胞仪鉴定转染细胞中存在CD40L表达。结论成功地建立人CD40L表达体系，为构建以人CD40L为组件的抗肿瘤疫苗的临床研究奠定基础。

Abstract

Objective To construct human eukaryotic expression system expressing T cell surface molecule CD40L and identify the expression product in SCCL cell line h446. Methods The expression vector pcDNA3.1-CD40L was constructed by recombinant DNA technology

then transfected into h446 cell line of SCCL by lipidosome methods

and the expressed membrane protein was identified by flow cytometry. Results The pcDNA3.1(+)-CD40L was constructed successfully and the coding region was inserted into the vector correctly. The membrane protein was observed by flow cytometry on transfected h446 cell line of SCCL. Conclusion Human CD40L was expressed successfully in eukaryotic system

which is very important in the anti-tumor vaccination study with a recombinant human CD40L in clinic.

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