结构关系-猫眼课题宝

结构关系

批准号：

3811195

负责人：

R Q HU

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：

至

来源：

https://reporter.nih.gov/project-details/3811195

关键词：

Escherichia coli gel electrophoresis immunoprecipitation interferon inducers interferons molecular cloning mutant polymerase chain reaction protein engineering protein structure function site directed mutagenesis transposon /insertion element

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英文摘要

It has been shown that deletion of residue -1 to 7 or 9 residues of rhu IFN -gamma results in the loss of biological activity (H. Hogrefe et al ref. J. Biol. Chem. 21, 12179-86, 1989). Due to the limited amount of material that can be isolated by enzymatic modification, we are using a protein engineering approach to study the relationship between the structure and function of human IFN-gamma. The human IFN-gamma gene has been cloned into PGEM 2 vector and PKK223-3 IFN-gamma gene expressing vector has been constructed. Immunoprecipitation assay and SDS PAGE revealed that cells induced by IPTG produced human IFN-gamma in E. coli JM105. Using PCR and oligonucleotide directed mutagenesis, several mutant genes which contain truncate or replacement at N-terminal will be obtained. Some mutant genes will be cloned into the expression vector, and characterized.

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