Expression and characterization of human TALL-1 in <em>Pichia Pastoris</em>

WANG Meng; YANG Yuan; YANG Zheng-rong; LI Guang-kuo; LIU Xiao-hong; SU Yong-ping; CHENG Tian-min

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Expression and characterization of human TALL-1 in Pichia Pastoris

更新时间：2025-01-15

- Expression and characterization of human TALL-1 in Pichia Pastoris
- Journal of Army Medical University Vol. 29, Issue 6, Pages: 494-497(2007)
- 作者机构：
  
  第三军医大学军事预防医学院防原医学教研室，全军复合伤研究所，创伤、烧伤与复合伤国家重点实验室
- 作者简介：
- 基金信息：
- DOI：
  CLC： R394.3;R379;R394-33
- Published：2007，
  
  Published Online：30 March 2007，
- 稿件说明：
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WANG MENG, YANG YUAN, YANG ZHENG-RONG, et al. Expression and characterization of human TALL-1 in Pichia Pastoris. [J]. Journal of army medical university, 2007, 29(6): 494-497.
DOI：

WANG MENG, YANG YUAN, YANG ZHENG-RONG, et al. Expression and characterization of human TALL-1 in Pichia Pastoris. [J]. Journal of army medical university, 2007, 29(6): 494-497. DOI：

摘要

目的研究在毕赤酵母（

Pichia Pastoris

）系统中人TALL-1基因的分泌表达，获得具有生物活性的重组TALL-1。方法将人TALL-1基因插入含AOX1启动子和α分泌信号肽序列的酵母表达载体pPIC9K中，用

Sac

Ⅰ将重组质粒线性化，电转化

Pichia Pastoris

GS115感受态细胞，筛选阳性整合子并进行甲醇诱导表达，采用MTT对表达产物进行初步的生物活性检测。结果经过诱导表达条件的优化，人TALL-1在酵母培养基BMMY中实现了分泌表达；免疫印迹和酶联免疫检测结果显示，重组产物可以结合其特异抗体；生物学活性检测证实其可抑制Hela细胞增殖。结论在毕赤酵母系统中实现了人TALL-1的分泌表达，具有生物学活性的重组TALL-1分子可以用于进一步的结构与功能研究。

Abstract

Objective To study the secretion expression of human TALL-1 cDNA in

Pichia Pa

storis and to obtain recombinant human TALL-1 of good biological activity. Methods The human TALL-1 gene was cloned into yeast expression vector pPIC9K containing AOXI promoter and the sequences of secreting signal peptides. Recombinant plasmid was linearized by

Sac

Ⅰ and transformed into

Pichia Pastoris

GS115 competent cells. Positive integrated clones were screened out and cultured in flasks

and hTALL-1 was expressed under the induction of 1% methanol. Biological activities were analyzed with ELISA and MTT assay. Results Human TALL-1 gene was expr

essed in BMMY after optimization of inducing expression condition. The recombinant could combine with specific antibody and the expressed product could inhibit the proliferation of Hela cells. Conclusion The secretion expression of human TALL-1 was successfully achieved in

Pichia Pastoris

expression system. The recombinant human TALL-1 could be used for further study on its structure and functions.

关键词

TALL-1分泌表达毕赤酵母

Keywords

TALL-1secretion expressionPichia pastoris

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Expression and characterization of human TALL-1 in Pichia Pastoris

DOI：

摘要

Abstract

关键词

Keywords

references