人干扰素α物种O和F的CDNA克隆和表达-猫眼课题宝

人干扰素α物种O和F的CDNA克隆和表达

批准号：

3792470

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/3792470

关键词：

SDS polyacrylamide gel electrophoresis complementary DNA gene expression human genetic material tag interferons molecular cloning protein sequence western blottings

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英文摘要

An IFN-alpha2 cDNA and the monoclonal antibodies NK2 and 4F2 were used as probes to screen a human Namalwa cDNA library to find the cDNA coding for component 0.21 positive clones which strongly hybridize with IFN-alpha2 cDNA were obtained. Due to the strong sequence similarity of the human IFN-alpha family, a specific oligonucleotide was applied to the 21 positive clones for component o to find cDNA coding for component o. We have performed the partial amino acid sequence of IFN-alpha component o isolated from human IFN-alpha results from sequence analysis of component o revealed that it is very similar to IFN-alphaF. In order to know the relationship between component o and IFN-alphaF, we cloned the gene for IFN-alpha into several expression vectors. Analysis by western blotting, SDS PAGE and antiviral activity revealed IFN-alphaF gene was expressed well in an pKK 223-3 expression system. We are now preparing sufficient quantities of IFN-alphaF to perform further studies.

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