用mRNA差异显示法比较SHR和WKY大鼠肾脏组织的基因表达

THE GENE EXPRESSION OF KIDNEY IN SPONTANEOUSLY HYPERTENSIVE RAT AND WKY RAT BY mRNA DIFFERENTIAL DISPLAY

目的用ｍＲＮＡ差异显示技术观察ＳＨＲ与ＷＫＹ大鼠肾脏组织在ｍＲＮＡ表达水平上的差异。方法取１２周龄的ＳＨＲ及ＷＫＹ大鼠，提取肾脏总ＲＮＡ，用含１２个寡聚核苷酸的Ｔ１１Ａ或Ｔ１１Ｇ引物进行逆转录，用Ｔ１１Ａ、ＡＰ１～３，Ｔ１１Ｇ、ＡＰ６，Ｔ１１Ｇ、ＡＰ７～８引物对逆转录产物进行多聚酶链式反应，反应产物进行聚丙烯酰胺凝胶电泳及放射自显影。结果每一种引物组合在单一电泳泳道上可显示７０～１００条基因表达条带，其中绝大多数代表肾脏基因表达的条带是相同的，但其中显示的４条ＳＨＲ大鼠肾脏基因表达的条带与相对应的ＷＫＹ大鼠肾脏基因表达的条带出现了差异。讨论ｍＲＮＡ差异显示技术对于快速筛选ＳＨＲ及ＷＫＹ大鼠肾脏组织的差异表达基因是适用的。

URPOSE To observe the gene expression of the kidney in spontaneously hypertensive rat(SHR) and wistar kyoto(WKY) rat by means of mRNA differential display.METHODS The technique of mRNA differential display is able to show the change of gene expression at mRNA level. The total RNA of the kidney extracted from the SHR and WKY rat at the age of 12 weeks was reverse transcribed (RT) to cDNA using anchored oligo(dT) primer T11A or T11G. PCR was performed using the above cDNA from the RT and arbitrary primer (AP). The primer combination in the PCR process included T11A and AP1～3, T11G and AP6, and T11G and AP8. The reaction samples were resolved on a polyacrylamide gel, and then, the autoradiography was performed.RESULTS More than 70 to 100 bands were visualized in each lane of the electrophrosis. The pattern of gene expression was same on the whole, but 4 cDNA fragments of the SHR were different from those of the WKY.CONCLUSIONS The mRNA differential display method is suitable for the rapid screening of differentially expressed genes in the kidney of the SHR and WKY rats.