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两种不同小鼠干扰素-β mRNA 的部分 cDNA 拷贝的分子克隆

Molecular cloning of partial cDNA copies of two distinct mouse IFN-beta mRNAs.

作者信息

Skup D, Windass J D, Sor F, George H, Williams B R, Fukuhara H, De Maeyer-Guignard J, De Maeyer E

出版信息

Nucleic Acids Res. 1982 May 25;10(10):3069-84. doi: 10.1093/nar/10.10.3069.

Abstract

Two cDNA libraries were constructed, using respectively the 12S and the 16S sucrose gradient fractions of polysomal poly (A)+ RNA from mouse C243 cells induced with Newcastle disease virus. Screening of a part of both libraries by mRNA selection hybridization assays revealed the presence of two plasmids hybridizing to an mRNA, whose translation product was characterized as mouse IFN-beta. Blot analysis of RNA indicated that mRNA hybridizing to the DNA from both plasmids could be detected in induced but not in uninduced C243 cells. The two cDNA inserts did not cross hybridize and had distinct restriction maps. Sequencing revealed that both inserts represented the end of the coding region and the entire 3' non coding region of two district mRNAs. Although different, the putative 39 AA and 65 AA carboxy termini of both Mu IFN-beta s display some homology to human IFN-beta 1. Thus there are at least two different murine IFN-beta genes.

摘要

构建了两个cDNA文库,分别使用新城疫病毒诱导的小鼠C243细胞的多聚核糖体多聚(A)+ RNA的12S和16S蔗糖梯度级分。通过mRNA选择杂交分析对两个文库的一部分进行筛选,结果显示存在两个与一种mRNA杂交的质粒,其翻译产物被鉴定为小鼠IFN-β。RNA的印迹分析表明,在诱导的C243细胞中可以检测到与来自两个质粒的DNA杂交的mRNA,而在未诱导的C243细胞中则检测不到。这两个cDNA插入片段不发生交叉杂交,并且具有不同的限制性图谱。测序显示,这两个插入片段均代表两个不同mRNA的编码区末端和整个3'非编码区。尽管不同,但两种小鼠IFN-β的推定39个氨基酸和65个氨基酸的羧基末端与人类IFN-β1具有一些同源性。因此,至少存在两个不同的小鼠IFN-β基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3efd/320691/ca29e00405c2/nar00379-0055-a.jpg

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