CDNA CLONING AND EXPRESSION OF HUMAN IFN-ALPHA SPECIES O AND F

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: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 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

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.

使用IFN-α 2 cDNA和单克隆抗体NK 2和4F 2作为对照。 探针筛选人Namalwa cDNA文库，以找到编码 组分0.21与IFN-α 2强杂交的阳性克隆 获得cDNA。 由于人类的强序列相似性， IFN-α家族，一种特异性寡核苷酸应用于21 组分o的阳性克隆以找到编码组分o的cDNA。 我们对IFN-α组分的部分氨基酸序列进行了测定， 〇从人IFN-α分离的结果来自以下的序列分析： 组分o显示其与IFN-α F非常相似。 为了 知道组分o和IFN-α F之间的关系，我们克隆了 将IFN-α基因导入几种表达载体中。 西方分析 杂交、SDS-PAGE和抗病毒活性检测结果表明，IFN-α F基因在大肠杆菌中表达， 在pKK 223-3表达系统中表达良好。 我们现在正准备 足够量的IFN-α F以进行进一步的研究。