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克隆人类性染色体共有的一个表达基因。

Cloning an expressed gene shared by the human sex chromosomes.

作者信息

Darling S M, Banting G S, Pym B, Wolfe J, Goodfellow P N

出版信息

Proc Natl Acad Sci U S A. 1986 Jan;83(1):135-9. doi: 10.1073/pnas.83.1.135.

Abstract

The existence of genes shared by mammalian sex chromosomes has been predicted on both evolutionary and functional grounds. However, the only experimental evidence for such genes in humans is the cell-surface antigen encoded by loci on the X and Y chromosomes (MIC2X and MIC2Y, respectively), which is recognized by the monoclonal antibody 12E7. Using the bacteriophage lambda gt11 expression system in Escherichia coli and immunoscreening techniques, we have isolated a cDNA clone whose primary product is recognized by 12E7. Southern blot analysis using somatic cell hybrids containing only the human X or Y chromosomes shows that the sequences reacting with the cDNA clone are localized to the sex chromosomes. In addition, the clone hybridizes to DNAs isolated from mouse cells that have been transfected with human DNA and selected for 12E7 expression on the fluorescence-activated cell sorter. We conclude that the cDNA clone encodes the 12E7 antigen, which is the primary product of the MIC2 loci. The clone was used to explore sequence homology between MIC2X and MIC2Y; these loci are closely related, if not identical.

摘要

基于进化和功能方面的原因,人们预测哺乳动物性染色体存在共享基因。然而,在人类中,此类基因的唯一实验证据是由X和Y染色体上的基因座(分别为MIC2X和MIC2Y)编码的细胞表面抗原,该抗原可被单克隆抗体12E7识别。利用大肠杆菌中的噬菌体λgt11表达系统和免疫筛选技术,我们分离出了一个cDNA克隆,其主要产物可被12E7识别。使用仅含人类X或Y染色体的体细胞杂种进行Southern印迹分析表明,与该cDNA克隆反应的序列定位于性染色体上。此外,该克隆与从已转染人类DNA并在荧光激活细胞分选仪上筛选出12E7表达的小鼠细胞中分离出的DNA杂交。我们得出结论,该cDNA克隆编码12E7抗原,它是MIC2基因座的主要产物。该克隆用于探索MIC2X和MIC2Y之间的序列同源性;这些基因座即使不完全相同,也是密切相关的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fbbd/322806/c98ec94adfb4/pnas00305-0150-a.jpg

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