Application of mRNA Differential Display to the Identification of Genes Related to Embryonic Development  

mRNA差异显示技术在动物早期胚胎发育相关基因研究中的应用

作　　者：李书鸿[1] 韩文[1] 马海飞[1] 

机构地区：[1]中国科学院发育生物学研究所

出　　处：《Developmental and Reproductive Biology》1997年第2期67-75,共9页发育与生殖生物学报（英文版）

摘　　要：mRNA differential display was established by Liang and Pardee in 1992 for the purpose of displaying the mRNA differences between two tissues. The early embryonic development in animals is primarily controlled by the maternal RNAs stored in egg. These mRNAs are being degraded as the development proceeds. In some animals, such as fish and amphibian, new transcripts do not appear until the midblastula stage (midblastula transition, MBT). In other animals, for example in mouse, the zygotic genes are expressed during very early stages of development. The mRNA programmed synthesis and degradation during embryonic development controls the cell differentiation, germlayer formation and pattern formation. All these mRNA changes could be displayed side by side as cDNA band differences by mRNA differential display and the genes corresponding to these differential mRNAs could thus be obtained.动物从卵裂开始的早期胚胎发育始终伴随着ｍＲＮＡ不断的合成与降解。最初受着受精卵内母体ｍＲＮＡ的控制。当这些ｍＲＮＡ在发育过程中逐渐被降解时，有些动物（如小鼠）在比较早的时期已有新基因的转录；而有些动物（如鱼类、两栖类）迟到囊胚中期以后，合子核的基因才开始转录新的ｍＲＮＡ。ｍＲＮＡ这种有规律的代谢活动控制着细胞的分化、胚层的形成以及模式形成（ｐａｔｅｒｎｆｏｒｍａｔｉｏｎ）。所有这些ｍＲＮＡ水平上的变化都可以用ｍＲＮＡ差异显示法（ｍＲＮＡｄｉｆｅｒｅｎｔｉａｌｄｉｓｐｌａｙ）检测出来并进而获得与早期胚胎发育有关的基因。ｍＲＮＡ差异显示法是由Ｌｉａｎｇ和Ｐａｒｄｅｅ于１９９２年建立起来的，用于显示两种不同组织之间ｍＲＮＡ差异的方法。基于绝大多数ｍＲＮＡ有一个ｐｏｌｙ（Ａ）尾巴，选用一个较特殊的３’端引物─５’Ｔ１１ＭＮ３’，其中Ｍ可以是ｄＡＴＰ，ｄＣＴＰ，ｄＧＴＰ之一，而Ｎ可以是ｄＡＴＰ，ｄＴＴＰ，ｄＣＴＰ，ｄＧＴＰ之一，进行逆转录时，１／１２的ｍＲＮＡ可被逆转录为ｃＤＮＡ。这种ｃＤＮＡ第一链被用来ＰＣＲ扩增，所使用的３’端引物与逆转录引物相同，而５’端引物为１０个碱基的一段寡核苷酸，这类任意（ａｒｂｉｔｒａｒ?

关 键 词：mRNA differential display embryonic development GENE 

分 类 号：Q344.5[生物学—遗传学] Q954.4