昆明小鼠早期胚胎PFK基因转录产物的检测

摘要

Inner and outer primer pair were designed and synthesizedaccording to the cDNA sequence of liver type phosphofructokinase (PFK) of the mouse. A nested reverse transcriptase-polymerase chain reaction (Nested RT-PCR) was used to determine the transcript pattern of mRNA encoding mouse PFK throughout 1-，2-，4-，8-cell and morulae embryos. The results showed that 1-cell to morulae mouse embryos mRNA could be amplified to give an expected band using the inner primer pair on the templates provided by the products of the first PCR using the outer primer pair. These observations indicated that the transcripts of PFK gene existed and that the PFK of early mouse embryos is of the liver type. Glycolysis and tricarboxylic acid cycle (TCA) appear to be the important mode of glucose metabolism in 1-cell to morulae mouse embryos.%根据小鼠肝脏型6-磷酸果糖激酶(PFK)的cDNA序列设计合成内外2对引物，以巢式RT-PCR方法检测昆明小鼠早期胚胎是否转录出PFK的mRNA.结果发现，1-细胞至桑椹期胚胎的mRNA均能以外引物扩增产物为模板通过内引物扩增出1条特异性目的片段.表明昆明小鼠1-、2-、4-、8-细胞至桑椹期胚胎的PFK以肝脏型的形式出现，其PFK基因已转录，糖酵解及三羧酸循环可能是早期胚胎葡萄糖代谢的主要方式.