用聚合酶链反应制备抗端粒酶蛋白重链和轻链可变区基因

张徽; 张波; 韩继生; 侯琳

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用聚合酶链反应制备抗端粒酶蛋白重链和轻链可变区基因

更新时间：2024-01-17

- 用聚合酶链反应制备抗端粒酶蛋白重链和轻链可变区基因
- Preparing heavy and light chain variable region genes for anti- telomerase protein hTERT by the polymerase chain reaction
- 重庆医科大学学报 2001年第2期页码：132-134
- 作者机构：
  
  1. 重庆医科大学基础医学院病理学教研室,重庆,400016
  2. 北京医科大学病理学教研室,北京,100083
- 作者简介：
- 基金信息：
- DOI：
  中图分类号： R392-33
- 纸质出版日期：2001，
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[1]张徽,张波,韩继生,侯琳.用聚合酶链反应制备抗端粒酶蛋白重链和轻链可变区基因[J].重庆医科大学学报,2001(02):132-134+138.

Preparing heavy and light chain variable region genes for anti- telomerase protein hTERT by the polymerase chain reaction[J]. Journal of Chongqing Medical University, 2001,(2):132-134.
[1]张徽,张波,韩继生,侯琳.用聚合酶链反应制备抗端粒酶蛋白重链和轻链可变区基因[J].重庆医科大学学报,2001(02):132-134+138. DOI：

Preparing heavy and light chain variable region genes for anti- telomerase protein hTERT by the polymerase chain reaction[J]. Journal of Chongqing Medical University, 2001,(2):132-134. DOI：

摘要

目的：为了制备抗端粒酶蛋白hTERT可变区基因。方法：采用Ni-NTA树脂层析法从PBKTRT转染的 E.coli JM109阳性菌中分离纯化端粒酶蛋白hTERT，并免疫小鼠。从免疫的小鼠脾脏中提取淋巴细胞中总RNA，逆转录成cDNA，以逆转录合成的cDNA第一条链为模板，分别用VH和VL引物，通过PCR进行扩增。结果：PCR扩增反应产生350bp大小的抗hTERT重链和轻链可变区基因。VH和VL基因片段的获得为进一步制备抗hTERT单功能区抗体（dABs）和单链抗体ScFv奠定了实验基础。结论：提示该方法具有稳定性好，易于重复，扩增后VH和VL量较高的优点。

Abstract

Objective: To make variable region genes against telomerase protein hTERT. Methods: Telomerase protein hTERT was purified from E.coli JM 109 harboring pBKTRT by Ni-NTA resin chromatograph and immunized Balb/c mice. Total RNA of lymphocytes were extracted from spleens of immunized mice and converted to cDNA by RT-PCR. First-strant cDNA was used as a template

heavy and light chain variable region genes for anti-telomerase protein hTERT were amplified individually with VH primers and VL primers using polymerase chain reaction. Results:The heavy and light chain amplification reaction yielded a fragment approximately 350 base pairs in length. These fragment could provide possibility for preparing single-chain antibodies（ScFv）against telomerase. Conclusion: Results suggest that the method have better stability and easy repetition

and could get higher quantity of VH and VL.

关键词

端粒酶重链可变区轻链可变区聚合酶链反应

Keywords

telomeraseHeavy chain variable regionLight chain variable regionPolymerase chain reaction

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