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EXPRESSION AND ISOLATION OF A FUNCTIONAL IFN ALPHA RECEPTOR FROM INCLUSION BODIES

EXPRESSION AND ISOLATION OF A FUNCTIONAL IFN ALPHA RECEPTOR FROM INCLUSION BODIES
功能性 IFN α 受体的表达和从包涵体中的分离
批准号:
3792480
负责人:
N Y NGUYEN
金额:
$0.0万
依托单位:
--
依托单位国家:
美国
项目类别:
财政年份:
--
资助国家:
美国
项目状态:
未结题
起止时间:

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中文摘要
翻译
编码IFN-α的整个胞外区的基因 已将受体(缺少信号肽)克隆到质粒pGEX-2 T中 转化大肠大肠杆菌DH 5作为融合蛋白与细菌 谷胱甘肽-S-转移酶 一种编码SEQ ID N 0:28-437的氨基酸的DNA片段, 将受体蛋白在EcoRI处插入载体pGEX-2 T中, BamHJ网站。 受体下游的终止密码子 序列被添加。 IPTG诱导表达,融合蛋白 通过Sepharyl PS-300凝胶层析部分纯化蛋白质 随后进行谷胱甘肽琼脂糖亲和层析。 工作正在 蛋白酶切割后回收非融合受体的进展 凝血酶,并启动物理化学和生物学研究 重组蛋白的表征;抗 受体将被产生,并将用于纯化天然蛋白质 从哺乳动物细胞,并将作为一个探针,研究机制 干扰素-α及其受体参与的信号转导。
英文摘要
The gene coding for the entire extracellular domain of the IFN-alpha receptor (lacking the signal peptide) has been cloned in plasmid pGEX-2T and expressed in E. coli DH5 as a fusion protein with bacterial glutathione-S-transferase. A DNA fragment encoding amino acids 28-437 of the receptor protein was inserted into vector pGEX-2T at the EcoRI and BamHJ sites. A stop codon immediately downstream of the receptor sequence was added. Expression was induced by IPTG and the fusion protein was partially purified by gel chromatography on Sepharyl P S-300 followed by glutathione agarose affinity chromatography. Work is in progress to recover the non-fused receptor after-protease cleavage by thrombin, and to initiate studies on the physico chemical and biological characterization of the recombinant protein; antisera against the receptor will be produced and will be used to purify the natural protein from mammalian cells, and will serve as a probe to study the mechanisms of signal transduction involving interferon-alpha and its receptor(s).
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STRUCTURAL AND BIOLOGICAL CHARACTERIZATION OF A RHIFN ALPHA RECEPTOR
  • 批准号:
    3748189
  • 项目类别:
  • 资助金额:
    $0.0万
  • 财政年份:
    --
  • 负责人:
    N Y NGUYEN
  • 依托单位:
    --
STRUCTURAL - BIOLOGICAL CHARACTERIZATION OF RECOMBINANT HUMAN IFN ALPHA RECEPTOR
  • 批准号:
    3770350
  • 项目类别:
  • 资助金额:
    $0.0万
  • 财政年份:
    --
  • 负责人:
    N Y NGUYEN
  • 依托单位:
    --
ISOLATION OF BIOLOGICALLY ACTIVE CYTOKINES BY NON-DENATURING PAGE
  • 批准号:
    3792462
  • 项目类别:
  • 资助金额:
    $0.0万
  • 财政年份:
    --
  • 负责人:
    N Y NGUYEN
  • 依托单位:
    --
ISOLATION OF BIOLOGICALLY ACTIVE CYTOKINES BY NON-DENATURAING PAGE
  • 批准号:
    3811202
  • 项目类别:
  • 资助金额:
    $0.0万
  • 财政年份:
    --
  • 负责人:
    N Y NGUYEN
  • 依托单位:
    --
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