Firsova N V, Markintantova Iu V, Smirnova Iu A, Panova I G, Sukhikh G T, Zinov'eva R D, Mitashov V I
Izv Akad Nauk Ser Biol. 2008 Mar-Apr(2):134-8.
The goal of this study was the search for and structure-function analysis of the regulatory genes specific for pluripotent embryonic stem cells (ESCs). This was the first study in which PCR was used to obtain DNA fragments with primers constructed on the basis of OCT4 and NANOG mRNA. cDNA synthesized on mRNA isolated from human embryonic eye in the 9.5th week of development was used as a template in PCR analysis. PCR fragment DNA was sequenced. A comparative analysis of the nucleotide sequences demonstrated their 100% homology with the OCT4-pg1 retrogene and NANOG gene. Expression of the genes of interest was reliably detected in the cornea, crystalline lens, retina, and eye tunics in the 10.5th week of development. The nuclear localization of the products of the NANOG gene and OCT4-pg1 retrogene indicates that these proteins are classified with transcription factors. The role of the OCT4-pg1 retrogene and NANOG gene in self-renewal and differentiation of pluripotent cells in a developing eye is discussed.
本研究的目的是寻找多能胚胎干细胞(ESC)特异性的调控基因并进行结构功能分析。这是第一项使用PCR基于OCT4和NANOG mRNA构建引物来获取DNA片段的研究。从发育第9.5周的人胚胎眼中分离的mRNA合成的cDNA用作PCR分析的模板。对PCR片段DNA进行测序。核苷酸序列的比较分析表明它们与OCT4-pg1反转录基因和NANOG基因具有100%的同源性。在发育第10.5周时,在角膜、晶状体、视网膜和眼膜中可靠地检测到了感兴趣基因的表达。NANOG基因和OCT4-pg1反转录基因产物的核定位表明这些蛋白质属于转录因子。讨论了OCT4-pg1反转录基因和NANOG基因在发育中的眼睛中多能细胞自我更新和分化中的作用。